cmo specialize to build better biologic...

The Science & Business of Biopharmaceuticals

INTERNATIONAL

www.biopharminternational.com

INTERNATIONAL

Bio

Ph

arm

Intern

atio

nal

SEP

TEM

BER 2

015

Sin

gle

-Use

Syste

ms I P

rote

in C

hara

cte

rizatio

n I P

roce

ss Co

ntro

ls V

olu

me 2

8 N

um

ber 9

September 2015

Volume 28 Number 9

CMOs SPECIALIZE TO BUILD BETTER BIOLOGIC DRUGS

OUTSOURCING

CMOs CONCERNED

WITH COST OF

SINGLE-USE EQUIPMENT

DOWNSTREAM

PROCESSING

CELL HARVESTING

SYSTEMS

TROUBLESHOOTING

STABILITY

TESTING

IN BIOPHARMA

ES670678_BP0915_cv1.pgs 09.09.2015 19:43 ADV blackyellowmagentacyan

CAPTURE

WITH NEW SELECTIVITY.(GAIN BOTH YIELD AND PURITY.)

See how Nuvia™ cPrime™ Mixed-Mode Resin

can transform your capture step.

Visit our booth and request a resin sample

BioProduction, Dublin, Ireland BioProcess International, Boston, MA

October 14–15, 2015 October 26–29, 2015

ES660009_BP0915_CVTP1_FP.pgs 08.20.2015 19:04 ADV blackyellowmagentacyan

Af�nity capture is usually not an option

for the downstream processing of

recombinant proteins, viruses, and second-

generation mAb-like proteins. Process

developers seek high recovery and purity

during initial capture. Using a mixed-mode

resin for capture enhances selectivity and

delivers increased total protein recovery

and purity.

We demonstrated mass capture of a

recombinant human adenovirus using

the hydrophobic cation exchange

Nuvia™ cPrime™ Mixed-Mode Resin

(Figure 1). This step of the puri�cation

achieved a tenfold reduction in processing

volume and a reduction in feedstream

contaminants.

This ef�cient two-step puri�cation process,

Nuvia cPrime Mixed-Mode Resin followed

by Nuvia Q Anion Exchange Resin,

provided approximately 54% overall

recovery of virus particles (Table 1).

Learn more at bio-rad.com/info/capture

NUVIA cPRIME RESIN IN A cGMP-READY PURIFICATION PROCESS FOR ADENOVIRUS PURIFICATION

AU

240 250 260 270 280 290

Time, min

1.2

0

60

0

mS

/cm

100 mM

NaOH

(strip)

Product collection

75 mM Tris, 525 mM NaCl,

pH 8.5 (elution)

Fig. 1. Initial capture of recombinant virus. OD 260 (—);

OD 280 (—); conductivity ( —). AU, absorbance units.

Table 1. Viral particle recovery and impurity clearance.

25 mM histidine, pH 6.0 (wash)

Sample

Total virus

(x1011 particles)

Impurity levels

(ng/1010 particles)

DNA HCP

Bulk harvest 30.6 3,144 n/d

Nuclease-treated harvest 31.8 30 3,020

Nuvia cPrime eluate 18.4 n/d 58

Nuvia Q eluate 16.4 <0.02 2

HCP, host cell protein; n/d, not determined.

ES660010_BP0915_CVTP2_FP.pgs 08.20.2015 19:04 ADV blackyellowmagentacyan


INTERNATIONAL

www.biopharminternational.com

INTERNATIONAL

Bio

Ph

arm

Intern

atio

nal

Sep

tem

ber 2

015

Sin

gle

-Use

Syste

ms I p

rote

in C

hara

cte

rizatio

n I p

roce

ss Co

ntro

ls V

olu

me 2

8 N

um

ber 9

September 2015

Volume 28 Number 9

CMOs speCialize tO build better biOlOgiC drugs

OutsOurCing

CMOs CONCERNEd

WITh COsT Of

sINgLE-UsE EqUIpMENT

dOwnstreaM

prOCessing

CELL hARvEsTINg

sysTEMs

trOubleshOOting

sTAbILITy

TEsTINg

IN bIOphARMA

ES670678_BP0915_cv1.pgs 09.09.2015 19:43 ADV blackyellowmagentacyan

www.tosohbioscience.com

Tosoh Bioscience and TOYOPEARL are registered trademarks of Tosoh Corporation.

Every mAb is unique.Every mAb is unique.

Your Protein A should be as well.

TOSOH BIOSCIENCE LLC • Customer service: 866-527-3587 • Technical service: 800-366-4875, option #3

TOYOPEARL® AF-rProtein A HC-650FHigh Capacity Protein A Resin for Monoclonal Antibody Purifi cation

0

10

20

30

40

50

60

70

80

2 3.5 5

Residence time (minutes)

DB

C f

or

IgG

(g

/L)

1 g/L

5 g/L

10 g/LResin: TOYOPEARL AF-rProtein A HC-650F

Column size: 5 mm ID × 5 cm

Mobile phase: 0.02 mol/L sodium phosphate, 0.15 mol/L NaCl, pH 7.4

Residence time: 2, 3.5, 5 min

Detection: UV @ 280 nm (10% breakthrough)

Sample: human IgG @ 1, 5, 10 g/L in mobile phase

ES668476_BP0915_CV2_FP.pgs 09.03.2015 02:31 ADV blackyellowmagentacyan

INTERNATIONAL


EDITORIALEditorial Director Rita Peters [email protected] Editor Agnes Shanley [email protected] Editor Susan Haigney [email protected] Editor Randi Hernandez [email protected] Science Editor Adeline Siew, PhD [email protected] Director Dan Ward [email protected] Editors Jill Wechsler, Jim Miller, Eric Langer, Anurag Rathore, Jerold Martin, Simon Chalk, and Cynthia A. Challener, PhD Correspondent Sean Milmo (Europe, [email protected]) ADVERTISING

Publisher Mike Tracey [email protected]/Mid-West Sales Manager Steve Hermer [email protected] Coast Sales Manager Scott Vail [email protected] Sales Manager Chris Lawson [email protected] Sales Manager Wayne Blow [email protected] List Rentals Tamara Phillips [email protected] 877-652-5295 ext. 121/ [email protected] Outside US, UK, direct dial: 281-419-5725. Ext. 121 PRODUCTION Production Manager Jesse Singer [email protected] AUDIENCE DEVELOPmENT Audience Development Rochelle Ballou [email protected]

UBm LIfE SCIENCES

Joe Loggia, Chief Executive Officer Tom Ehardt, Executive Vice-President, Life Sciences Georgiann DeCenzo, Executive Vice-President Chris DeMoulin, Executive Vice-President Rebecca Evangelou, Executive Vice-President, Business Systems Julie Molleston, Executive Vice-President, Human Resources Mike Alic, Executive Vice-President, Strategy & Business Development Tracy Harris, Sr Vice-President Dave Esola, Vice-President, General Manager Pharm/Science Group Michael Bernstein, Vice-President, Legal Francis Heid, Vice-President, Media Operations Adele Hartwick, Vice-President, Treasurer & Controller

UBm AmERICAS

Simon Foster, Chief Executive Officer Brian Field, Chief Operating Officer Margaret Kohler, Chief Financial Officer

UBm PLC

Tim Cobbold, Chief Executive Officer Andrew Crow, Group Operations Director Robert Gray, Chief Financial Officer Dame Helen, Alexander Chairman

© 2015 Advanstar Communications Inc. All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical including by photocopy, recording, or information storage and retrieval without permission in writing from the publisher. Authorization to photocopy items for internal/educational or personal use, or the internal/educational or personal use of specific clients is granted by Advanstar Communications Inc. for libraries and other users registered with the Copyright Clearance Center, 222 Rosewood Dr. Danvers, MA 01923, 978-750-8400 fax 978-646-8700 or visit http://www.copyright.com online. For uses beyond those listed above, please direct your written request to Permission Dept. fax 440-756-5255 or email: [email protected].

UBM Life Sciences provides certain customer contact data (such as customers’ names, addresses, phone numbers, and e-mail addresses) to third parties who wish to promote relevant products, services, and other opportunities that may be of interest to you. If you do not want UBM Life Sciences to make your contact information available to third parties for marketing purposes, simply call toll-free 866-529-2922 between the hours of 7:30 a.m. and 5 p.m. CST and a customer service representative will assist you in removing your name from UBM Life Sciences’ lists. Outside the U.S., please phone 218-740-6477.

BioPharm International does not verify any claims or other information appearing in any of the advertisements contained in the publication, and cannot take responsibility for any losses or other damages incurred by readers in reliance of such content.

BioPharm International welcomes unsolicited articles, manuscripts, photographs, illustrations, and other materials but cannot be held responsible for their safekeeping or return.

To subscribe, call toll-free 888-527-7008. Outside the U.S. call 218-740-6477.

EDITORIAL ADVISORY BOARDBioPharm International’s Editorial Advisory Board comprises distinguished specialists involved in the biologic manufacture of therapeutic drugs, diagnostics, and vaccines. Members serve as a sounding board for the editors and advise them on biotechnology trends, identify potential authors, and review manuscripts submitted for publication.

K. A. Ajit-Simh President, Shiba Associates

Rory Budihandojo Director, Quality and EHS Audit

Boehringer-Ingelheim

Edward G. Calamai Managing Partner

Pharmaceutical Manufacturing

and Compliance Associates, LLC

Suggy S. Chrai President and CEO

The Chrai Associates

Leonard J. Goren Global Leader, Human Identity

Division, GE Healthcare

Uwe Gottschalk Vice-President,

Chief Technology Officer,

Pharma/Biotech

Lonza AG

Fiona M. Greer Global Director,

BioPharma Services Development

SGS Life Science Services

Rajesh K. Gupta Vaccinnologist and Microbiologist

Jean F. Huxsoll Senior Director, Quality

Product Supply Biotech

Bayer Healthcare Pharmaceuticals

Denny Kraichely Associate Director

Johnson & Johnson

Stephan O. Krause Principal Scientist, Analytical

Biochemistry, MedImmune, Inc.

Steven S. Kuwahara Principal Consultant

GXP BioTechnology LLC

Eric S. Langer President and Managing Partner

BioPlan Associates, Inc.

Howard L. Levine President

BioProcess Technology Consultants

Herb Lutz Principal Consulting Engineer

EMD Millipore Corporation

Jerold Martin Independent Consultant

Hans-Peter Meyer Lecturer, University of Applied Sciences

and Arts Western Switzerland,

Institute of Life Technologies.

K. John Morrow President, Newport Biotech

David Radspinner Global Head of Sales—Bioproduction

Thermo Fisher Scientific

Tom Ransohoff Vice-President and Senior Consultant

BioProcess Technology Consultants

Anurag Rathore Biotech CMC Consultant

Faculty Member, Indian Institute of

Technology

Susan J. Schniepp Fellow

Regulatory Compliance Associates, Inc.

Tim Schofield Managing Director

Arlenda, USA

Paula Shadle Principal Consultant,

Shadle Consulting

Alexander F. Sito President,

BioValidation

Michiel E. Ultee Principal

Ulteemit BioConsulting

Thomas J. Vanden Boom Vice-President, Global Biologics R&D

Hospira, Inc.

Krish Venkat CSO

AnVen Research

Steven Walfish Principal Statistician

BD

Gary Walsh Professor

Department of Chemical and

Environmental Sciences and Materials

and Surface Science Institute

University of Limerick, Ireland

ES670766_BP0915_003.pgs 09.09.2015 22:54 ADV blackyellowmagentacyan

4 BioPharm International www.biopharminternational.com September 2015

Contents

INTERNATIONAL

BioPharm International integrates the science and business of

biopharmaceutical research, development, and manufacturing. We provide practical,

peer-reviewed technical solutions to enable biopharmaceutical professionals

to perform their jobs more effectively.

COLUMNS AND DEPARTMENTS

BioPharm International ISSN 1542-166X (print); ISSN 1939-1862 (digital) is published monthly by UBM Life Sciences 131 W. First Street, Duluth, MN 55802-2065. Subscription rates: $76 for one year in the United States and Possessions; $103 for one year in Canada and Mexico; all other countries $146 for one year. Single copies (prepaid only): $8 in the United States; $10 all other countries. Back issues, if available: $21 in the United States, $26 all other countries. Add $6.75 per order for shipping and handling. Periodicals postage paid at Duluth, MN 55806, and additional mailing offices. Postmaster Please send address changes to BioPharm International, PO Box 6128, Duluth, MN 55806-6128, USA. PUBLICATIONS MAIL AGREEMENT NO. 40612608, Return Undeliverable Canadian Addresses to: IMEX Global Solutions, P. O. Box 25542, London, ON N6C 6B2, CANADA. Canadian GST number: R-124213133RT001. Printed in U.S.A.

BioPharm International is selectively abstracted or indexed in: • Biological Sciences Database (Cambridge Scientifc Abstracts) • Biotechnology and Bioengineering Database (Cambridge Scientifc Abstracts) • Biotechnology Citation Index (ISI/Thomson Scientifc) • Chemical Abstracts (CAS) • Science Citation Index Expanded (ISI/Thomson Scientifc) • Web of Science (ISI/Thomson Scientifc)

6 Guest Editorial Ensuring data integrity involves effort on an individual and global basis. Richard M. Johnson

8 Global News

10 Regulatory Beat Legislation to streamline drug development may get tangled up in user fee negotiations and drug pricing battles.Jill Wechsler

14 Perspectives on Outsourcing Suppliers indicate prices for single-use equipment are likely to increase. Eric Langer

18 Industry Insider FDA’s proposed guidance for quality metrics raises questions about quantifying the tangibles and intangibles of quality culture. Andrew Harrison and Susan Schniepp

52 Troubleshooting Three case studies illustrate some analytical methods important for stability testing. Stella-Christiana Chotou

55 Product Spotlight

56 New Technology Showcase

57 Ad Index

58 Vaccines Development Update

Contract Biomanufacturing Firms Become More SpecializedRandi Hernandez

Focusing on niche and specialty service

offerings gives contract biomanufacturing

organizations an opportunity to

differentiate in a crowded market. 22

High Titers and Perfusion Processes Challenge Cell Harvesting SystemsCynthia A. Challener

New single-use technologies and other

filtration systems are beginning to address

cost, throughput, and manufacturing

footprint demands. 28

Best Practices in Qualification of Single-Use SystemsWeibing Ding

The author discusses the current best

practices in technical qualification of

single-use systems. 32

Enhancing Protein Binding Studies with a Light-Scattering ToolkitDaniel Some

Light-scattering techniques are useful

for interaction studies when traditional

methods of analysis do not suffice. 40

Utilizing Run Rules for Effective Monitoring in ManufacturingAaron Spence

To enable efficient monitoring

systems, life-science companies

need to effectively apply run rules. 48

Volume 28 Number 9 September 2015

Dieter Spannknebel/Stocktrek Images/Getty Images; Dan Ward


www.gelifesciences.com/BioProcessGE, GE monogram, and Xcellerex are trademarks of General Electric Company. © 2014–2015 General Electric Company. First published Apr. 2014. GE Healthcare Bio-Sciences AB, Björkgatan 30, 751 84 Uppsala, Sweden.

29120140 AC 08/2015

The Xcellerex™ XDR-50 MO fermentor system is purpose-

built for microbial cultures and delivers performance you

can trust under tough conditions. With a robust design,

the single-use technology enables rapid batch-to-batch

changeover for increased process effi ciency and fl exibility.

When it comes to increasing facility utilization, we are a

partner like no other.

Increased fl exibility in microbial processes. Our single-use fermentor system.

upstream downstream single-use services

ES668427_BP0915_005_FP.pgs 09.03.2015 02:27 ADV blackyellowmagentacyan


Guest Editorial

Ensuring data

integrity involves

effort on

an individual

and global basis.

Data Integrity: Getting Back to Basics

I have been involved in the healthcare products industry for all of my profes-

sional life, more than 35 years in varied environments and operations in the

United States, Europe, Asia, Australia, and Latin America. During that time,

I have seen many changes in the manufacture and control of pharmaceutical

and biopharmaceutical products.

Some changes are inherent in the newer products that have been introduced,

but many of these changes have been technological: the advent of computer-

ized systems; instruments and sensors to monitor and control machine func-

tion; increasingly sophisticated software; and the ability to communicate data

via wire and wirelessly around the world.

One of the ways that we have applied these technologies is through the

generation of volumes of data that were previously either not accessible or

impractical to analyze. It is through the generation of data, and the review

of these data by knowledgeable persons in comparison to specifications and

standards, that we base our quality decisions. Even the least sophisticated

operation today employs electronic systems for many applications: inventory

control, analytical data capture and analysis, equipment control, and report-

ing. As these changes have occurred, there have been evolving changes in the

regulatory focus—from quality control and an emphasis on testing quality—to

quality assurance and process validation—to quality by design and lifecycle

management.

The human elementBut while electronic systems are becoming more widely used and generating

ever larger amounts of data, it is also true that all of our operations depend

on a network of people to perform a wide variety of functions, from material

manufacturing, transportation, finished-product manufacturing, testing, dis-

tribution, regulatory filings, and controlling records that involve a combina-

tion of both electronic and paper-based records.

The global elementIn addition, there is a growing globalization of the business and the depen-

dence on partners from outside the primary organization. Activities that used

to be done within a single organization are more likely today to include a net-

work of separate organizations that are interdependent on goods, services, and

data exchange. Many of the healthcare products used by patients around the

world involve manufacturers and suppliers from multiple continents who com-

municate in multiple languages.

The data elementA basic principle of assuring the quality of healthcare products is the review

of data. Industry experts review data from their partners; independent quality

groups review manufacturing and testing data; and regulators who are respon-

sible for monitoring the products for the public review all data. The accuracy,

trustworthiness, and the integrity of that data must not be in question, or all

of the checks and balances, control measures, and quality agreements will not

be effective. This is equally true of the traditional paper-based records and

electronic data.

The recent emphasis on data integrity is not new, but it has never been more

crucial. This principle should remind us of a basic tenet: All relationships are

based on trust and evidence. If the integrity of one’s data is questionable, the

loss of trust will have severe consequences. We should all be committed to

reinforcing the importance of data integrity. ◆

Richard M. Johnson is the president and CEO of the

Parenteral Drug Association.


FDA Releases Guidance on

Biosimilar NomenclatureThe long-awaited draft guidance on biosimilar naming

has been posted by FDA, and within the document,

the agency suggests naming follow-on biologics by a

common nonproprietary name, followed by a suffix that

will identify the manufacturer of the particular biosimilar.

The naming scheme is consistent with the way the agency

labeled Zarxio (filgrastim-sndz), the first biosimilar

product to be approved by FDA. The naming convention

will be used for all future biosimilar products and will also

be applied retroactively, meaning the naming convention

will apply to biologic products (submitted for approval

through the 351[k] or 351[a] pathways) that are already on

the market.

The renaming of all biologic medications has a dual

purpose, says FDA: to facilitate the “routine” use of

suffixes for biologics, and to “avoid inaccurate perceptions

of the safety and efficacy of biological products based

on their licensure pathway.” In other words, FDA does

not want healthcare providers to judge a drug’s quality

based on its name. As such, FDA will use the same

nonproprietary name for biosimilars (i.e., the generic or

“core name”) plus a manufacturer-specific suffix; taken

together, the core name and suffix will produce a “proper

name”.

Although FDA did not provide a full rollout plan for

how all the existing biologics will be renamed, it did

say that it intends to “assign distinguishing suffixes

to a limited group of these products.” The originator

biologics that will get new designations first are those

reference products that will be susceptible to biosimilar

competition in the near future, and FDA is considering

a rulemaking action to implement the retrospective

naming of biologics. For future products, manufacturers

will propose suffixes for their products themselves in

application documents. It is still unclear if these suffixes

will be required to directly reference the manufacturer, as

in the case of Sandoz’s filgrastim-sndz, or if the suffix will

be chosen based on other parameters, but FDA said the

suffix should be unique, “devoid of meaning”, and should

not be promotional. For example, proposing adalimumab-

best as a proper name for Humira would likely be blocked

by the agency. The suffix also cannot be a commonly used

abbreviation in clinical practice, contain reference to a

drug core name, or be confusingly similar to the originator

product’s suffix (once assigned) or to a similar product’s

suffix.

Some biological products will be exempt from the

renaming initiative, says FDA; these include products

“for which a proper name is provided in the regulations”

and those medications that already have robust

pharmacovigilance programs in place. FDA attests that

current originator products will benefit from a suffix

title, as it will help with pharmacovigilance efforts. It’s

unclear how many originator biologics will actually be

renamed, as the directives in the FDA guidance are meant

as suggestions, not requirements, and it may be difficult

to assess which drugs already have “proper names” by FDA

standards and which ones do not.

Though FDA said the naming protocol is meant to

prevent inadvertent substitution among biologic products

that have not been determined to be interchangeable,

the agency did not say how it will label interchangeable

biologics and how these are to be differentiated among

the other products in the same class. All the agency

said was that it was “considering whether the suffix

should be unique or should be the same as the reference

product.” If an interchangeable product shared the

same suffix as the originator, however, the tracking of

a drug’s safety by its suffix would not be feasible, and

pharmacovigilance problems could persist. Determination

of interchangeability and the naming of interchangeable

products will be covered in a future guidance, and FDA

requested feedback on how to name interchangeable

products.

—Randi Hernandez

GSK to Divest Ofatumumab

to Novartis Pharma

GlaxoSmithKline plc announced on Aug. 21, 2015 an

agreement with Novartis Pharma AG, a subsidiary of

Novartis AG, to divest its rights in ofatumumab for

auto-immune indications, including multiple sclerosis

for up to $1 billion, plus royalties.

Earlier this year, Novartis Pharma acquired the

oncology indications for ofatumumab (Arzerra) as

part of a three-part transaction between GSK and

Novartis. After completion of the latest transaction,

Novartis Pharma will own rights to ofatumumab in

all indications.

The consideration payable by Novartis Pharma to

GSK comprises milestone payments of $300 million

payable at closing; $200 million payable subject to

the start of a phase III study in relapsing remitting

multiple sclerosis by Novartis; and further contingent

payments of up to $534 million payable on the

achievement of certain other development milestones.

Novartis Pharma will also pay royalties of up to

12% to GSK on any future net sales of ofatumumab in

auto-immune indications.

The transaction is expected to complete by the end

of 2015. Glo

be im

ag

e: C

ha

d B

ake

r/G

ett

y I

ma

ge

s; P

ho

to C

red

it:


Global News


Sent 9:51 AM

A 10% improvement in yield!?!

Yes on top of a 15% shorter run time!Lab Tech || Mark D. || 9:54 AM

© 2015 Hamilton Company. All rights reserved.

1-888-525-2123www.hamiltoncompany.com

Incyte Measures Viable Cell Density in Real Time

Incyte is insensitive to media changes, microcarriers,

dead cells and fl oating debris. It can be used to monitor

changes in cell physiology, cellular respiration, viral infection

timing, automated harvesting and much more.

Optimize Your Process

Learn more at www.ham-info.com/1024

PLATINUM

ELECTRODES

ELECTRIC

FIELDS

VIABLE

CELLS

POLARIZE

DEAD CELLS HAVE DAMAGED

MEMBRANES AND

DO NOT POLARIZE



Regulatory Beat

Vis

ion

so

fAm

eri

ca

/Jo

e S

oh

m/G

ett

y Im

ag

es

The later months of 2015 wi l l be

important in determining the fate of

a number of programs slated to shape

drug development and industry health in

the years ahead. Legislation to advance bio-

pharmaceutical R&D faces hurdles on Capitol

Hill, despite overwhelming House approval

in July 2015. Important user fee negotiations

between FDA and biopharmaceutical compa-

nies have begun, but tight timeframes and

competing demands will make it difficult to

resolve issues quickly. A mounting backlash

against high price tags on important new

drugs, moreover, may bolster opposition to

initiatives designed to speed new therapies to

market.

Mov ing the 21st Cent ur y Cures Ac t

through Congress in 2015 will be tough, as

the Senate seeks changes and legislators con-

tend with the usual impasse over the federal

budget for fiscal year 2016, as well as debate

over the Iran nuclear arms pact and major

trade agreements. Next year will be even

worse, as the presidential election campaign

heats up, coinciding for the first time with

the process for re-authorizing user-

fee agreements. With a new admin-

istration and a new Congress in

January 2017, FDA fees and legisla-

tion need to be ready for review by

both outgoing and incoming offi-

cials by the middle of next year.

ProsPects for “cures”Despite broad enthusiasm for the

Cures legislation, the bi-partisan

support that moved it through the

House may fall apart in the com-

ing months. The measure nearly

faltered due to opposit ion from

consu mer advocate s c la i m i ng

it would bring more unsafe drugs to mar-

ket, and from fiscal conservatives opposed

to increased mandatory spending for the

National Institutes of Health (NIH). But

House Energ y & Commerce Committee

Chairman Fred Upton (R-Mich) and his allies

garnered support from hundreds of patient

groups, research organizations, medical soci-

eties, and biopharmaceutical companies, all

eager to increase basic research at NIH and

to provide FDA with added resources. These

and other stakeholders applauded provisions

to more fully incorporate patient experi-

ences in considering a drug’s benefits and

risks, expedite new drug approvals through

biomarker qualification, streamline clinical

trials, encourage sponsors to use data from

clinical experience, and provide incentives

for developing antibiotics and treatments for

rare diseases (1).

The House measure now faces revisions

in the Senate, where there is talk of devel-

oping a much narrower bill by fall. Some

Senate Republicans strongly oppose NIH bud-

get increases, which would erode Democratic

support. If the Senate does enact a slimmer

bill, it would face difficult conference com-

mittee negotiations with the House, and the

process could extend well into next year.

focus on feesConsequently, many provisions of the Cures

legislation may end up in a broad FDA bill

to reauthorize user fees for drugs, generic

drugs, medical devices, and biosimilars by

summer 2017. FDA launched negotiations

in June 2015 for revising the Generic Drug

User Fee Act (GDUFA II) and in July 2015 for

medical devices and for prescription drugs

(PDUFA VI). Monthly discussions with indus-

try will begin in September 2015, along with

Manufacturers Face Key Policy and Regulatory ChallengesLegislation to streamline drug development may get tangled up in user fee negotiations and drug pricing battles.

Jill Wechsler is BioPharm

International’s Washington editor,

chevy chase, MD, 301.656.4634,

[email protected].


Eppendorf®, the Eppendorf logo, BioBlu®, epMotion®, and Vacufuge® are registered trademarks of Eppendorf AG, Germany.

U.S. Design Patents are listed on www.eppendorf.com/ip

Of ers may vary by country. All rights reserved, including graphics and images. Copyright © 2015 by Eppendorf AG.

More great of ers for your cell biology processes — now save up to 30 %!

Our new limited of ers enable you to:

> Convert your existing bioprocess

system to advanced single-use

technology: with BioBlu® Single-

use Vessels and adaptor kits

> Automate your pipetting: with

epMotion® 5070 and 5075 liquid

handling stations

> Concentrate your samples in a

quick, ef cient, and gentle manner

using the Concentrator plus*

(*U.S./CAN: Vacufuge® plus)

> Go green and save on ultra-low tem-

perature freezers! Choice of various

models between 101 L and 725 L

www.eppendorf.com/advantage

New Promotion Sept. 1 – Dec. 31, 2015

Take Good Care of Your Cells



regulatory Beatregulatory Beat

regular meetings for FDA to hear

the views of patients, consumers,

and health professionals.

The importance of user-fee

programs to FDA was evident at

the PDUFA meeting with both

act ing commissioner Stephen

Ost rof f and deput y commis -

sioner Rober t Cal i f f of fer ing

comments, along with Center for

Drug Evaluation and Research

(CDER) Director Janet Woodcock.

But fast agreements and leaner

authorizing measures won’t be

easy to achieve. Generic-drug

makers are unhappy with FDA’s

slow pace in meeting the goals

set by the init ial GDUFA pro-

gram, noting that timeframes for

reviewing abbreviated new drug

applications (ANDAs) actually

have gone up, and there has been

little gain in whittling down the

massive ANDA backlog. Slow FDA

approval of new generic drugs

only aggravates shortage situa-

tions and delays patient access

to important, affordable medi-

cines, industry reps observed at

the June 2015 meeting.

Generic-drug firms also com-

plained about r ising fees, par-

t icu la rly when FDA had not

spent a big chunk of the money

it already has collected. Small

companies want relief in facility

fees, especially companies seek-

ing their f irst approved prod-

uct. Manufacturers urged greater

t ransparency and better com-

munication on agency decisions,

notably about a stiffer “refuse to

accept” policy that permits the

agency to reject ANDAs lacking

key components.

At the PDUFA public meeting,

patient groups and professional

organizations offered a range

of initiatives meriting user fee

support: more biomarker vali-

dation, pediatric and neonatal

drug development, data transpar-

ency initiatives, and greater con-

sistency in review practices by

review divisions. A main theme

was expanding the use of “real-

world” evidence to accelerate

drug development. Greg Daniel

of the Brook ings Inst itut ion

highlighted strategies for tap-

ping clinical evidence to support

agency decisions and to docu-

ment product safety, while Allan

Coukell of the Pew Charitable

Trusts emphasized the value of

observational data from claims

databases.

FDA is expected to seek access

to some portion of drug applica-

tion fees during product devel-

opment, instead of waiting until

f iling. Biosimilar sponsors pay

a portion of the fee upfront to

support CDER’s time and invest-

ment in product development

meet ings, a process that has

expanded with the proliferation

of breakthrough drugs and other

cruc ia l medic ines. Current ly,

only biosimilar sponsors pay a

fee to FDA even if no applica-

tion is filed at the end, points

out Gillian Woollett of Avalere

Health, a model that FDA would

like to extend to other products.

Pricing PerilsUser-fee negotiations assume a

certain number of production

faci l it ies and applicat ions for

drugs and medical products each

year to generate expected fees.

But these project ions may be

tempered by a slowdown in bio-

pharma R&D and sales, as payers

and health organizations attack

companies for setting unaccept-

ably high prices, as seen with the

recent emergence of important

new treatments for hepatitis C.

Oncologists are livid over soar-

ing prices for new cancer drugs,

as seen in a July 2015 editorial

in the Mayo Clinic Proceedings

signed by 118 physicians that

urge action to halt the r ising

prices (2). The American Society

of Clinical Oncology has devel-

oped a Cancer Value Framework

to relate the value of a drug and

its price to clinical benefits and

toxicities, a strategy applauded

by pharmacy benefit managers.

FDA approval of new PCSK9

inhibitors to manage cholesterol,

which could be used by millions

of patients, has set off a cam-

paign by payers and insurers to

limit indications and prescrib-

ing. Praluent (alirocumab), devel-

oped by Sanofi and Regeneron

Pharmaceuticals, was launched

in Ju ly 2015 with a $14,600

list price per year—much more

than anticipated and way more

than currently avai lable cho-

lesterol meds, but with labeling

designed to limit use to seriously

ill patients. There is much hope

for new treatments to prevent

Alzheimer’s disease, but already

strong pushback against poten-

tially high costs.

The Institute for Clinical and

Economic Review (ICER) is devel-

oping an assessment of the value

of PCSK9 inhibitors as part of

its expanded program to pro-

vide independent reports on the

cost- and comparative-effective-

ness and budget impact of new

drugs where the value evidence

is controversial and where the

budget impact may be quite high,

as with treatments for cancer,

asthma, and diabetes. The esca-

lating focus on drug “value” will

be important to policies and pro-

grams for biopharma R&D, pro-

duction, and marketing.

references 1. Rules Committee Print 114-22 Text

Of H.R. 6, 21st Century Cures Act,

http://docs.house.gov/

billsthisweek/20150706/CPRT-114-

HPRT-RU00-HR6.pdf

2. Mayo Clinic, In Support of a Patient-

Driven Initiative and Petition to Lower

the High Price of Cancer Drugs, Mayo

Clinic Proceedings 90 (8) (August

2015), www.mayoclinicproceedings.

org/article/S0025-

6196%2815%2900430-9/fulltext ◆


Buffers Mixers Cell Culture Media Clarification Services

Introducing Mobius® 2000 LSingle-use Bioreactor.

Ease of use with an innovative twist.

The world’s first bioreactor with a pull-out drawer for quick,

easy loading. That’s just the first of many innovations:

- Self-inflating bag, with built in baffle for

homogeneous mixing, eliminates the need

for operator intervention during inflation.

- 5:1 turndown ratio provides the most

flexible seeding and growth strategy.

- Intuitive, simple-to-navigate software

reduces the learning curve for operators

and allows for easy integration into

your plant automation strategy.

Mobius® 2000 LSingle-use Bioreactor

EMD Millipore Corp. is a subsidiary of Merck KGaA, Darmstadt, Germany

EMD Millipore, the M logo and Mobius are registered trademarks of Merck KGaA, Darmstadt, Germany.© 2015 EMD Millipore Corporation, Billerica, MA USA. All rights reserved. AD1419EN00 Rev A. 07/2015

www.emdmillipore.com/mobiustwist

ES671382_BP0915_A13_FP.pgs 09.11.2015 21:08 ADV blackyellowmagentacyan


Perspectives on Outsourcing

Do

n F

arr

all/G

ett

y Im

ag

es

Contract manufacturing organizations

(CMOs) have been enthusiastic adopters

of single-use technologies in recent years,

lauding their range of benefits and attributing

significant improvements to their use. In fact,

according to the latest industry study from

BioPlan Associates (1), there appears to be one

aspect of single-use application that CMOs feel

is inhibiting adoption: The cost.

As part of the BioPlan study, factors that

may restr ict the use of disposables were

examined and general agreement was found

among both biomanufacturers and CMOs

when segmenting these two groups. The top

two concerns for each group, for example,

were “breakage of bags and loss of produc-

tion” and “leachables and extractables”, with

the former the most widely held among bio-

manufacturers and the latter most common

among CMOs.

Other common pain points for both groups

include material incompatibility with process

fluids, single-source issues, and the high cost

of disposables, with each of these a factor in

restricting adoption of single-use devices for

between 55–60% of respondents. Meanwhile,

CMOs (62.5%) were far more likely than bio-

therapeutic developers (40.4%) to cite the

lack of clear regulatory guidance

on leachables and extractables

(L&E) as a hindrance.

When the more than 200 sur-

vey respondents were asked to

ident i f y the most impor tant

reason for not increasing use of

disposable technologies, a new

ranking of obstacles was found,

as differences between developers

and CMOs emerged (see Figure 1).

For biotherapeutic developers,

the most crucial reasons restrict-

ing greater use of disposables are:

• “Breakage of bags and loss of production

material is a concern” (19% of biothera-

peutic developers)

• “For regulatory reasons, we can’t change

our current systems” (15.5%).

For CMOs, however, the top reasons given

were:

• The high cost of disposables (consum-

ables), at 26.7% of respondents

• Leachables and extractables are a concern

(20%).

Cost obstacles for CMOs partly exist because

these service suppliers need to justify their cost

to their clients. CMOs also have significantly

more in-use experience with these devices, so

cost rises to the top as other factors decrease in

importance.

One area that CMOs indicate has not been

worked out is leachables and extractables. For

example, 13% of CMOs worry for the lack of

regulatory guidance on L&E—noting this as

their top obstacle. In combination with other

concerns about L&E, these could be viewed

as the leading technical challenge to CMO

adoption.

It is interesting that single-use equipment is

generally accepted as providing cost savings, yet

the single most-cited factor restricting further

use by CMOs is cost.

Given that CMOs expect to increase their

budgets by as much as 6% for new technolo-

gies to improve upstream and downstream effi-

ciency—with single-use equipment presumably

a part of this spending—budgetary constraints

are not holding back single-use adoption.

Is sIngle-use too expensIve?The relatively rapid growth in use of single-

use equipment for early-stage (and increas-

ingly commercial stage) manufacturing may

suggest that costs are considered reasonable.

Yet as in-use experience grows, it is expected

that the high cost of some of these con-

CMOs Concerned With Cost of Single-Use EquipmentSuppliers indicate prices for single-use equipment are likely to increase.

Eric Langer is president of

Bioplan Associates,

tel. 301.921.5979,

[email protected].


September 2015 www.biopharminternational.com BioPharm International 15

perspectives on outsourcing

sumables will create friction and

push-back. As industry adoption

increases, single-use products in

biopharmaceutical manufacturing

will likely see the same levels of

pricing pressure as other similar

equipment. Vendors can expect

to see increasing price push-back

from their customers, and poten-

tially greater levels of competition.

sAtIsfActIon wIth vendorsThe influence of pricing as a selec-

tion criterion is unsurprising, of

course. Cost factors are right up

there with product quality, on-

schedule delivery, and the provi-

sion of L&E data that regulators will

accept. However, for this industry,

and most others, cost factors are a

commonly complained about issue.

Frankly, it’s rare to find buyers that

are particularly satisfied with the

cost of anything they buy.

The dissatisfaction with the cost

of single-use devices among buy-

ers of single-use devices is more

likely the result of buyers not per-

ceiving the full value that these

technologies can provide. This per-

ception is partly because this is a

relatively new segment and there

is little economic data, or in-use

experience and cost justifications

that are readily available. As more

data are developed, it is likely this

dissatisfaction gap will decrease.

In addition, as more fully inte-

grated single-use systems facilities

are brought online, the value of a

totally disposable operation will

begin to be realized.

In examination of the gap

between the importance of vari-

ous factors—including pricing and

the satisfaction end-users have

expressed regarding their single-

use vendors— approximately one in

five respondents (21.6%) reported

being “satisfied” or “very satisfied”

with single-use device costs. By

comparison, close to three-quarters

reported being satisfied with the

quality of the products (see Figure 2).

In any case, satisfaction with

single-use pricing is unlikely to

change anytime soon. Separately,

the BioPlan study also surveyed 164

suppliers to the biopharmaceutical

industry on issues associated with

industry growth. Suppliers reported

having increased their pricing for

disposable, single-use devices by

an average of 3.1% in 2014, about

the same average price increase as

the year before. Suppliers, however,

projected an average 3.8% increase

in pricing for disposable, single-use

devices this year, outstripping pro-

jected increases for other bioman-

ufacturing-related areas including

services, consumables, and instru-

ments. These results suggest that the

overall market for single-use equip-

ment is trending towards higher

prices, most likely due to consistently

high demand.

does cost reAlly mAtter to cmos?Single-use equipment costs are

unlikely to decrease and, as noted,

complaints about product costs

may be simply lack of understand-

Figure 1: Selected “Most Important Reasons” for not increasing use of

disposables, biotherapeutic developer vs. CMOs.

26.7%

0.0%

6.7%

0.0%

20.0%

13.3%

13.3%

4.8%

7.1%

7.1%

10.7%

10.7%

15.5%

19.0%

High cost of disposables (consumables)

We do not want to become vendor-dependent (single-source issues)

Limited scalability over a broad range (such as 2 L to 2,000 L)

We have already invested in equipment for current system

Leachables and extractables are a concern

For regulatory reasons, we can't change our current systems

Breakage of bags and loss of production material is a concern

Biotherapeutic Developer CMO

Which is the MOST important reason for not increasing use of disposable technologies?

“Biotherapeutic Developers vs CMO”

Source: 12th Annual Report and Survey of Biopharmaceutical Manufacturing, April 2015, www.bioplanassociates.com/12th

Figure 2: Percentage point gap between importance of single-use systems (SUS)

product attributes and level of satisfaction, select responses.

46.00%

46.20%

52.80%

54.40%

58.40%

Performing leaches/extractables testing using my current systems

Deliver on schedule

Providing leachables andextractables data that regulators will accept

Cost of product

Willingness to accept and fx integrity (leakage) problems

Source: 12th Annual Report and Survey of Biopharmaceutical Manufacturing, April 2015, www.bioplanassociates.com/12th

SUS Vendor Service Delivery GapsRelative Gap Between

Satisfaction with Vendor Delivery & Attribute’s Importance



perspectives on outsourcing

ing of the economics associated

with an effective application of the

technology. The market demand

continues, though, and CMOs give

a long list of reasons for increasing

use of disposables, from faster cam-

paign turnaround time (85.7%)

and reduced time to get the facility

up and running (75%) to decreased

risk of product cross-contamina-

tion (68.8%) and the elimination

of cleaning requirements (58.8%).

So given the benefits, does

the cost of these devices matter?

Indeed, while CMOs are more apt

than biotherapeutic developers

to find fault with single-use pric-

ing, they’re also more likely in

general to recognize the benefits

they afford.

While CMOs may ultimately

want the cost of single-use prod-

ucts to come down, high costs have

not stunted their adoption rates

to a considerable degree. Although

CMOs can pass the costs of such

single-use devices and operations

on to their clients, allowing CMOs

to reduce the potential need for

capital expenditures, there is an

additional factor noted that sug-

gests single-use systems device

costs are not fully relevant—almost

9 in 10 (86.4%) of CMOs surveyed

report at least “some” productivity

improvements due to the use of

disposable/single-use devices.

Thus, CMOs appear to be

addressing their most crucial trend

in global biomanufacturing—the

need to improve productivity and

efficiency, by adopting single-

use devices. So complaints about

costs may be a bit of disingenuous,

especially in relation to the pro-

ductivity benefits reportedly being

delivered.

reference 1. BioPlan Associates, 12th Annual

Report and Survey of

Biopharmaceutical Manufacturing

Capacity and Production (Rockville,

MD, April 2015), www.

bioplanassociates.com/12th. ◆

while cmos may ulti-

mately want the cost

of single-use products

to come down, high

costs have not stunted

their adoption rates to

a considerable degree.

VISIT OUR WEBSITE TO SUBSCRIBE FOR FREE TODAY! www.BioPharmInternational.com

www.twitter.com/BioPharmIntl

www.linkedin.com/groups/BioPharm-International-4027200

BIOPHARM

INTERNATIONALFIRST LOOK

Preview the latest

issue of BioPharm

International with quick

links to online content,

expanded coverage,

and the digital edition

of the magazine.

BIOPHARM’S SCIENCE & BUSINESS

BULLETIN

Monthly newsletter that

features in-depth articles

and timely news on

technologies for biotech

drug development and

manufacturing, and

regulatory developments,

as well as coverage of

deals, alliances, new

products, people moves,

and industry events.

BIOPHARM’S E-APPLICATION

NOTE ALERT

Receive monthly

application notes

categorized by

technique.

Sign up for BioPharm International FREE eNewsletters


[email protected] | 1 800 441 4225 | emergentcontractmanufacturing.com

Aseptic Fill/Finish

Pre-Clinical Development

BDSManufacture

Clinical & Commerical

Lyo CycleDevelopment

Emergent BioSolutions has a proven

track record as a world-class provider

of contract manufacturing services for both

bulk drug substances and sterile injectable

drug products. Our team has successfully

launched 20 commercial products and

developed/manufactured over 200

clinical candidates for our clients.

Emerging Capability

State-of-the art fexible single-use facility

enables turnkey upstream and downstream

solutions for clinical and commercial scale

biopharmaceutical drug development.

Emergent Contract Manufacturing:

Enhancing Life in Every Single Dose



Industry Insider

12

3re

nd

er/

E+

/Ge

tty Im

ag

es

The Metrics of Quality CultureFDA’s proposed guidance for quality metrics raises questions about quantifying the tangibles and intangibles of quality culture.

The long awaited, anxiously anticipated

FDA guidance on qual ity metr ics

was finally distributed for comment

on July 28, 2015. The official title of this

guidance for industry is Request for Quality

Met r ics, Guidance for Indust ry (1) and its

potential release has been looming on the

horizon since 2012.

The intent of FDA to establish quality

metrics first emerged in 2012 when Congress

passed the Food and Drug Administration

S a fe t y a nd I n novat ion Ac t ( F DA SI A)

enhancing FDA’s capability to proactively

react to, prevent, and alleviate drug short-

ages. Specifically, Title VII Section 705 of

the Act states FDA “shall inspect establish-

ments described in paragraph [1] that are

engaged in the manufacture, preparation,

propagation, compounding, or processing of

a drug or drugs (referred to in this subsec-

tion as ‘drug establishments’) in accordance

with a risk-based schedule established by

the Secretary.” Section 706 of the same act

allows FDA to request certain information

from companies in advance of or in lieu of

inspections by stating, “Any records or other

information that the Secretary may inspect

under this section from a person that owns

or operates an establishment that is engaged

in the manufacture, preparation, propaga-

tion, compounding, or processing of a drug

shall, upon the request of the Secretary, be

provided to the Secretary by such person, in

advance of or in lieu of an inspection …” (2).

In the Feb. 12, 2013 Federal Register Notice (3),

FDA asked the industry to “assist the Food

and Drug Administration in draft-

ing a strategic plan on drug short-

ages as required by the Food and

Drug Administration Safety and

Innovation Act …” This notice asked

a series of thought-provoking ques-

tions including “What metrics do manufac-

turers currently use to monitor production

quality?” and “How frequently would such

metrics need to be updated to be meaningful?”

After a few years of actively engaging and lis-

tening to industry in a variety of venues, this

new guideline has finally been released.

The metrics proposed in the guideline are

not new to industry. Many of them are cur-

rently being used by companies to internally

measure performance. In some cases, the speci-

fied metrics are also reported to the agency

via the annual report or are contained in the

annual product review. The bio/pharma indus-

try needs to review these metrics and ensure

they will provide meaningful data while avoid-

ing unintended consequences.

Defining quality cultureThe underlying and understated tenet used

to determine a company’s well-being is a

measure of their quality culture. The culture

of a company dictates the veracity of their

metrics. The best way to ensure the data

reported have merit is to assess the quality

culture of the submitting organization. It

is in this area that the new guidance lacks

clarity. The guidance leaves the opportunity

open to establish quality-culture metrics by

stating, “these metrics are not intended to

be an all-inclusive set of the quality metrics

that FDA could consider useful to assess a

product and manufacturer’s state of qual-

ity. For example, senior management com-

mitment to quality is an important factor

in evaluating the overall health of the PQS

[pharmaceutical quality system] and qual-

ity culture” (1) and “…the Agency is com-

mitted to a dialog with industry to consider

benchmarks and standards that could pro-

vide acceptable metr ics that specif ically

demonstrate senior management’s commit-

Andrew Harrison is chief regulatory

affairs officer and general

counsel, and Susan Schniepp is

distinguished fellow, both with

regulatory compliance associates.


That saved the life of my child.

To the breakthrough

trial

That was transported

direct

Who sourced the drug

Who contacted the expert

You are the person

Running a clinical trial requires close

collaboration between many different people.

But however complex the process, we never

lose sight of our objective – to help you bring lifesaving

medicines to market.

Make the connection with Clinigen CTS:

Email: [email protected]

Web: www.clinigengroup.com/clinical-trial-services

Trust our chain reaction



industry insider

ment to a culture of quality …” (1).

This commitment to establishing

quality culture metrics is further

evidenced by the section in the

guideline titled “Optional Metrics

Related to Quality Culture and

Process Capability/Performance”.

In this section, FDA “acknowledges

the importance of quality culture

to the overall state of quality of the

product, process, and commitment

to quality” (1).

Metrics relateD to quality cultureFDA proposes three voluntary met-

rics to try to get at the elusive qual-

ity culture. The first optional metric

proposed is intended to measure

senior management engagement

by assessing whether the head of

the quality unit and the head of

the operations unit have signed

the annual product review (APR) or

product quality review (PQR). The

second optional metric proposed

is corrective action and preventive

action (CAPA) effectiveness. The

measurement for this metric is to

indicate the percentage of correc-

tive actions that required retrain-

ing of personnel, the assumption

being that the root cause of the orig-

inal deviation (real or due to insuf-

ficient analysis) was determined to

be insufficient or ineffectual train-

ing. The third proposed metric is

intended to measure a firm’s process

capabilities through a series of three

questions. The real question should

be if these three optional metrics,

taken together, shed any light on the

quality culture.

Management engagement

Achieving a quality culture requires

management and employees to

establish an environment where

responsibility, accountability, and

reliability are paramount, and to

understand the role each person

performs in delivering a high-qual-

ity product to the customer and

sustaining that performance on a

continual basis. Management must

educate employees and provide the

tools and environment where they

can perform their functions in an

atmosphere that encourages excel-

lence and continuous improvement.

Assigning the head of quality and

the head of operations the task of

signing the APR or the PQR does

not ensure management engage-

ment nor does it mean that the

quality culture is lacking. It is up

to an organization to establish the

appropriate level of responsibility

and signing authority for APR and

PQRs. It is up to senior manage-

ment to provide the people charged

with these activities the necessary

resources to complete the task in a

timely manner with the expectation

that they will be held accountable

for the contents.

Retraining personnel

The second optional quality cul-

ture metric is specific to CAPA. The

proposed metric is to report the

percentages of corrective actions

involving the retraining of person-

nel. Without context supporting

the retraining of personnel, this

metric does not offer insight into

the true culture of an organization.

It could be argued that any CAPA

that results in a reduction or elim-

ination of a recurring deviation

would require an element of train-

ing personnel. In fact, retraining of

personnel on the CAPA issue, how

it was solved, and how to imple-

ment the necessary change is evi-

dence of management engagement.

It should be expected that a major-

ity of CAPAs involve some retrain-

ing of personnel.

Critical quality attributes

The third quality-culture optional

metrics involves trying to use criti-

cal quality attributes (CQA) as a

key indicator of a quality culture.

Of the three optional metrics pro-

posed, this one does provide some

measurement of the existence of a

quality culture. On the surface, the

questions just seem to be a regurgita-

tion of information contained in the

APR or PQR. Upon closer evaluation,

however, it is clear that FDA is try-

ing to measure whether a company

drives for continuous improvement

through their review and assess-

ment of threshold levels established

with CQAs. Companies that have

established CQAs and linked them

to a requirement to issue a CAPA

when they exceed the established

threshold levels have demonstrated

a commitment to continuous

improvement. Continuous improve-

ment programs are, in fact, reliable

indicators of the presence of a qual-

ity culture.

Measuring intangiblesThe establishment of simple qual-

ity metrics that not only measure

the quality of the product but also

reflect the quality culture of an

organization is required to assist

FDA in establishing a risk-based

audit program. The problem is that

it is difficult to measure something

as intangible as culture with cold,

hard data. The remaining ques-

When choosing a

metric, it is important

that the architects

of the metric are

aware of unintended

consequences that

may inadvertently

drive negative

behavior.



industry insider

tion is: If taken together, are the

three proposed optional metrics

indicative of a quality culture? The

answer is, maybe.

Careful thought and consider-

ation should be exercised when

determining what to measure, how

often to measure, how to interpret

and communicate the data, and

what the expectation is for using

the data to drive positive change.

Management needs to be cognizant

of the fact that whatever metrics are

reported, they must be developed,

evolved, and adjusted over time to

maximize their impact on driving

positive change. When choosing

a metric, it is important that the

architects of the metric are aware of

unintended consequences that may

inadvertently drive negative behav-

ior. Management attempting to

incentivize achievement of the goal

such as offering a financial award

if the goal is achieved, may lead

to inappropriate behaviors that do

not address the real issue. In these

cases, it is generally not the met-

ric that will drive the behavior but

rather use of behavioral rewards.

Reward for achievement rather

than analysis of the real underlying

causes will not lead to sustainable

positive change. When managed

properly, metrics are an important

tool to help drive positive change

and quality process improvements.

Upon observation, an unhealthy

quality culture is easy to identify.

People in a poor culture do not

understand their job and its impor-

tance to the business. They often

appear stressed, and they hide their

mistakes or blame others for their

errors. There is no evidence of team-

work. People work in silos and rarely,

if ever, seek input or advice from

others. Metrics that could potentially

be used to measure a poor culture

include a large employee turnover,

an overabundance of deviations

attributed to human error, and

lack of pride in the performance of

employees’ jobs.

In contrast, a robust, healthy

quality culture can be evidenced by

alignment of goals between qual-

ity and operations, self-sustained

work teams that focus on continual

improvement, and employees who

incorporate quality into their jobs

on a daily basis. They are not afraid

to speak up and offer suggestions

for improvement to their colleagues.

People understand the importance

of their jobas and respect each other

and their management. This culture

welcomes inspections and views

these inspections as another tool

to use in their continual improve-

ment initiatives. Metrics that could

potentially be used to measure a

healthy quality culture include a

small employee turnover, deviations

that identify a root cause other than

human error, and pride in the per-

formance of their jobs.

conclusionWhen establishing a metrics pro-

gram, companies should evaluate

numerous data input points includ-

ing, but not limited to, product-qual-

ity attributes, manufacturing site

performance, people metrics, and

quality-system metrics. For product-

quality metrics, companies should

consider reporting on batch-specific

data such as trending drug product,

drug substance, and stability test

results against customer complaint

rates. Indirect product-quality met-

rics could include environmental

monitoring, water trend results, and

yield rates. When establishing site

metrics, the company could look at

inspection history including internal

audit findings and maintenance his-

tory such as equipment age versus

defect-failure rates. People metrics

should consider ongoing job-spe-

cific training and education, skills

and experience assessments, and

employee turnover rate by job func-

tion and site. Quality systems met-

rics might look at change control,

investigation root-cause trends, and

release-testing cycle times.

There is no set requirement on

which metrics a company should

track to measure their overall perfor-

mance. Each company should deter-

mine which metrics to track based

on their operations, number of facili-

ties they operate and where they are

located, what types of products they

manufacture, and what type of cul-

ture exists in their places of business.

The metrics chosen must be

meaningful and written to provide

a clear analysis of ongoing activities.

It is important for operations and

quality to agree on the metrics and

how to report them to management

to avoid overreaction to the data. It

is not sufficient to simply report the

data. The interpretation of the data

is of crucial importance because it

may include a root-cause analysis of

its own.

references 1. FDA, Request for Quality Metrics

Guidance for Industry (Rockville, MD,

July 2015).

2. The Food Drug Administration Safety

and Innovation Act, Pub. L. 112-144,

126 Stat. 993 (2012).

3. FDA, “Food and Drug Administration

Drug Shortages Task Force and

Strategic Plan; Request for Comments”

Federal Register Feb. 12, 2013) online,

www.gpo.gov/fdsys/pkg/FR-2013-02-

12/html/2013-03198.htm, accessed

Aug. 17, 2015. ◆

When managed

properly, metrics

are an important

tool to help drive

positive change

and quality process

improvements.



Die

ter S

pannkneb

el/S

tocktr

ek

Imag

es/

Gett

y Im

ag

es;

Dan W

ard

Assessing the capabilities of

a contract manufacturing

o r g a n i z a t io n (C M O) i s

a col laborat ive proces s ,

involving numerous communications

be t ween t he b iopha r maceut ica l

company and the CMO. Choosing a

manufacturing partner solely based

on scale, on prior experience with

the CMO client, or on the supposed

reputat ion of the CMO may have

been fruitful strategies in the past—

but as the requirements for purity,

quality, and comparability become

more precise, the select ion of an

appropriate business partner for the

manufacture of a sensitive biologic

product can sometimes prove to be

as challenging as the bioprocessing

techniques themselves.

Whi le the demand for outsourc-

ing se r v ices i s s tead i ly inc reas -

ing, say Eric Langer and Ron Rader

of BioPlan Associates, sizing up the

various partner options and capa-

bilit ies among CMOs and contract

development and manufac tur ing

organizations (CDMOs) can be diffi-

cult. Experts within the outsourcing

industry are unclear about the total

number of CMOs/CDMOs. Even gaug-

ing the percentage of CMOS/CDMOs

focused specifically on biomanufac-

turing is a challenge. In June 2014,

15 CMOs/CDMOs united to become

part of the Pharma & Biopharma

Outsourcing Association (PBOA)—a

contract biomanufacturing firms become more specialized

Randi Hernandez

Focusing on niche and specialty

service offerings gives contract

biomanufacturing organizations an

opportunity to differentiate in a crowded market.

contract biomanufacturing


October 15-16, 2015

Puerto Rico Convention CenterSan Juan, Puerto Rico

WWW.INTERPHEXPUERTORICO.COM/BIOPHARM

CO-LOCATED WITH:PREMIER SPONSOR: SHOW SPONSOR:

INTERPHEX Puerto Rico is the place to learn and experience the latest in pharmaceutical, biopharmaceutical and medical device Innovation, Technologies and Knowledge.

Enhanced Technical Education Program offering educational and training opportunities to increase productivity and improve effciency.

Effective and effcient way to solve your development and manufacturing challenges, deliver greater value, drive performance and identify partnering opportunities to meet your production goals and objectives.

Network with your industry colleagues as you interact with and procure the latest technology under one roof.

Bringing together Key Decision Makers like YOU with industry leading solutions and technologies exhibitors for pre-scheduled face-to-face meetings right on the show foor!

Learn about higher product quality with continuous manufacturing! Show Floor Tours, Demonstrations and Technical Presentations. Watch for additional details.



nonprofit trade association for

CMOs/CDMOs representing the

interests and concerns of the

industry on matters affecting

the development and/or manu-

facture of pharmaceutical prod-

ucts—but the organizations that

have joined the trade group (now

at 18 members) encompass out-

sourcing companies from both

pharma and biopharma. Led by

journalist-turned-advocate, Gil

Roth, who is president, the trade

group covers issues such as the

assignment of Gener ic Dr ug

User Fee Amendment (GDUFA)

fees for facility inspections, and

the quality agreements between

CMOs and their clients.

C M O s h a v e d i f f e r e n t

approaches to quality agreements,

and the PBOA hopes that the

FDA’s draft guidance on the topic

will help lead to harmonization

or a proliferation of best practices

in that area. In an adjacent area,

the association is working with

FDA to help clarify the agency’s

recent draft guidance, Request for

Quality Metrics (1). Roth spoke at

an FDA public meeting on the

draft guidance, where he pushed

for clarity on the role of CMOs in

providing quality metrics to help

assure product safety, without cre-

ating a huge compliance burden.

The PBOA is also concerned that

certain metrics could favor in-house

manufacturing over outsourcing,

and Roth hopes PBOA can ensure

that the CMO perspective is repre-

sented when FDA ultimately imple-

ments the quality metrics (2).

Langer and Rader est imate

that there are as many as 80

biologics CMOs, and approxi-

mate ly 20 of these ac t ua l ly

make biologics. The three com-

panies in the top tier—Lonza,

Boehringer Ingelheim (BI), and

Patheon—make up the major-

it y of the biolog ics market ,

accord ing to BioPlan. Other

industry insiders argue, how-

ever, that while Lonza, BI, and

Patheon are among the biggest

players in the f ield, Celltr ion

and Samsung have signif icant

capacity in mammalian cell cul-

ture—and Sandoz is likely the

biggest CMO for microbial bio-

logics. Two CMO players that

fo l low t he a fo r e me nt ione d

companies in terms of business

activities with biologics include

Fujifilm Diosynth Technologies

and CMC Biologics, followed by

approximately 10 other com-

panies that make up the rest at

clinical scale. Jon S. Gingrich,

manager of business develop-

ment at Avid Bioservices, says

that Avid’s planned commercial

facility expansion will “triple its

present commercial capacity.”

The remaining 60-plus biolog-

ics CMOs are “specialty” CMOs,

according to Langer and Rader,

many of which are local businesses.

In short, approximately 15–20% of

the biologics CMOs actually man-

ufacture drugs, but five times as

many CMO facilities provide ser-

vices that support the production

of large-molecule medications.

“The market for biolog ica l

CDMO services—both in drug

substance and drug product—

is certainly robust,” observes

Roth. Areas such as biosimilars

and antibody-drug conjugates

(ADCs) present huge opportuni-

ties for drug makers and CDMOs

alike, he notes, adding that these

fields require significant levels

of expert ise and capital—and

players lacking in these attr i-

butes wil l be kept out of the

game. “The biologic space isn’t

for the fainthearted, and there

are a range of CDMO models,”

says Roth. “This demonstrates

that there isn’t one clear ‘plan of

attack’ for services and technol-

ogy solutions providers.”

The successful CMOs are those

that have continuously diversi-

fied their service offerings, note

Langer and Rader, and those

that have specialized in a par-

ticular niche area in the process

stream outside of the basic ser-

vice offerings of typical CMOs.

Some examples of this variety—

outside of the typical CMO ser-

vice offerings, such as process

development—include exper-

tise in bioconjugation and ADCs

(SAFC, Cata lent, Lonza, and

Goodwin Biotechnology); Baxter

Biopharma Solutions’ proficiency

in formulation development and

lyophilization cycle development

for biopharmaceuticals; Althea’s

ability to create stable crystal-

line formulations for therapeu-

tic products in suspension and

its expertise in protein expres-

sion using a non-endotoxic bac-

terial platform; KBI Biopharma’s

expanded microbial development




and manufacturing services via

its acquisition of a Merck facil-

ity; Fujifilm Diosynth’s aptitude

in diverse gene therapy; WuXi

AppTec’s mammalian cell cul-

ture capabilities through the use

of disposable bioreactors and

its dedication to enhancing the

manufacture of cell therapeu-

t ics; Lonza’s exper ience with

viral gene and virally modified

cell products; and MicroProtein

Technologies’ proficiency in a

fu l ly automated, plate -based

prote i n produc t ion met hod

using Escher ichia coli . Langer

calls MicroProtein Technologies’

recombinant protein platform

“quite revolutionary”; the com-

pany touts itself as the “first com-

pany to produce recombinant

biologics on an industrial scale

without employing the conven-

tional fermentation process” (3).

Expectations of a CMO

handling unstable products

As it relates to the handling of

products made in living cells,

“a good CMO will be able to

support both development and

manufacturing programs with a

raft of orthogonal techniques to

analyze both in-process samples

and final purified material,” says

Daniel Smith, chief scientif ic

officer, Cobra Biologics. CMOs

i n t he b iolog ic space must

comprehend the factors affecting

product stabi l ity (parameters

such as temperature, pH, and

buf fer concentrat ion, among

others); have a good grasp on

p ote nt i a l p ro duc t s t r e s sor s

(shear force and pump type);

have informat ion on proven

hold t imes for intermediates

and final products; have a solid

s c ient i f ic u nde r s t a nd i ng of

protein degradation pathways;

a n d a c c e s s t o e x c e l l e n t

charac ter i zat ion methods to

me a su re pu r i t y a nd de te c t

aggregation.

Bioassays are still on top

Contract service providers are

expected to benef it f rom the

surge of interest in biosimilar

development, especially when

it comes to product character-

izat ion assays used to assess

comparability between a refer-

ence product and a biosimilar.

The demand for accurate and

sens it ive b ios im i la r compa-

rabil ity tools has prompted a

renewed interest in the use of

orthogonal techniques to char-

acterize elusive structural dif-

ferences between molecules (4).

Disciplined formulation prac-

tices for biosimilars are of para-

mount importance, says Wendy

Saffell-Clemmer, research and

development director at Baxter

BioPharma Solutions. With inno-

vator medications, the process

defines the product—whereas for

biosimilars, the process must be

designed to deliver the targeted

product, she adds. Thus, prod-

uct characterization through the

use of orthogonal bioanalytical

test methods can help provide

insight on whether the develop-

ment process has been appropri-

ately designed.

“FDA’s tota l ity of ev idence

approach has analytical char-

acterization at its foundation,

which will l ikely result in an

increase in analyt ical serv ice

requests during cell-line selec-

t ion, fermentat ion, and puri-

f icat ion deve lopment ,” says

Saffell-Clemmer. A recent survey

from BioPlan Associates found

that biomanufacturers rely pri-

marily on CMOs for analytical

test ing and for bioanaly t ica l

methods that measure the purity

or biological activity of a drug

product (5). Approximately 86%

of biologics manufacturers use

CMOs for analytical testing ser-

vices; a slightly lower number

compared to 89% in 2014 (6).

The other four activities mak-

ing the top five most commonly

outsourced serv ices in 2015,

according to the BioPlan sur-

vey, are the same as they were

in 2014: plant ma intenance

services, f ill/finish operations,

validation services, and toxic-

ity testing. Plant maintenance

services jumped from the fifth

most com mon ly out sou rced

activity in 2014 to the second

most commonly outsourced ser-

vice in 2015, perhaps indicating

that biopharmaceutical compa-

nies’ aging facilities may drive

businesses to outsource to CMOs

in lieu of investing in new equip-

ment for their own facilities. The

increased use of automation and

disposable technologies in asep-

tic processes are also expected to

drive business to CMOs, as many

CMOs have already invested in

the technology to do this work,

such as the purchase of large-

scale isolators for the preparation

of potent compounds (7).

An uptick in service requests

related to the development of

biosimilars has been observed

by many of the CMOs that spoke

to BioPharm International. These

services are primarily related to

analytical testing, with a special

interest in the understanding of

product glycosylation patterns,

notes Smith. Gingrich confirms

that his company has seen a

signif icant increase in glyco-

prof i l ing requests; this assay

is currently one of the most-

requested types at Avid.

O t he r a na ly t ic a l s e r v ice s

expected to gain popularity as

more biosimilar candidates flood

the pipeline include formula-

tion screenings, high-throughput

screenings, and quality and com-

parability studies, according to

Sébastien Ribault, PhD, director

of Provantage Biodevelopment/

e nd - to - e nd s e r v ice s , Me rc k





BioDevelopment, Millipore S.A.S.,

France. To f ind the opt imal

clone, comparative analytics for

biosimilars should ideally begin

as early in the process as cell-line

development, suggests Marion

Schrader, PhD, senior director of

marketing at Rentschler. Despite

the increased interest in biosimi-

lars, Schrader ment ions that

Rentschler expects “to see some

consolidation in the area, as not

all of the current clinical projects

will have commercial success.”

Other commonly outsourced

activities include: API manufac-

ture, cell-line development, cell

banking (master cell banks and

working cell banks), process devel-

opment, formulation develop-

ment, conjugation development,

media development, and routine

production processes. Activities

that present unique facility con-

cerns—such as the manufacture

of ADCs, cell and gene therapies,

compendial and stability test-

ing, or viral vaccines and vec-

tors—are usually outsourced as

well. Firelli Alonso-Caplen, PhD,

senior director of biotherapeutics

and vaccines outsourcing at Pfizer

Biotech, notes that keeping more

complex projects in house, how-

ever, helps her company in par-

ticular retain internal full-time

employees. She says these chal-

lenging projects “become motiva-

tional drivers for more innovation

for our own scientists.”

T he r e w a s no c on s e n s u s

among industry experts as to

whether viral clearance studies

are usually outsourced or not.

According to Gingr ich, most

CMOs do not have experience

working with viral banks and

many of them do not want to

have any viral material onsite.

On the other hand, P f i zer ’s

Alonso-Caplen says that v iral

vaccines and vectors pose a risk

to in-house biopharmaceutical

manufactur ing faci l it ies, and

such act iv it ies are, therefore,

likely be outsourced to CMOs.

While many activities are out-

sourced in a piecemeal fashion,

and the need for certain services

can vary widely depending on

a biopharmaceutical sponsor’s

existing core capabilities, there

is always the possibility that a

product sponsor could come to a

CMO with a completely defined

process and ask the CMO to

solely assume a production role.

The reason for this production-

driven request could be because

the pharma c l ient has over-

whelmed its in-house capacity,

or because it wants the CMO to

manufacture “a secondary supply

of clinical or commercial prod-

ucts for [the purpose of] risk mit-

igation,” explains Smith.

When outsourcers outsource

A CMO may decline a project

if the client requests fall out-

side of the scope of its capabili-

t ies. When a CMO suspects it

cannot complete certain tasks

as requested, it is important for

that company to “be upfront and

transparent with the client so

that [the client is] able to pursue

discussions with other CMOs,”

notes Jennifer Cannon, PhD,

senior director of commercial

strategy at Ajinomoto Althea.

Out-of-scope requests are one

of the “great chal lenges that

sharing project or partnership details

CMOs and their pharma clients are generally not allowed to disclose details of their partnerships, although the decision to disclose

information varies by company. Some see the value in public disclosures, says Jennifer Cannon, PhD, senior director of commercial

strategy at Ajinomoto Althea, whereas others want to keep their business dealings private. Daniel Smith, chief scientific officer,

Cobra Biologics, says pharma’s reticence to disclose partner/manufacturing details is often due to the competition to be first-to-

market with a product. Jon S. Gingrich, manager of business development at Avid Bioservices, says that its contracts in highly

competitive areas, such as biosimilars, explicitly state that information on the molecules being developed remain confidential. In

the quest to produce biosimilars of comparable quality and efficacy to innovator products, biosimilar manufacturers may comb

through patent documents to gain insight about compound formulation and process platforms used by branded drug makers. This

is why details surrounding the partnerships between CMOs and branded biologic manufacturers may be another information entry

point for biosimilars to gain a competitive advantage for comparability, and perhaps eventually, interchangeability.

Some clients give permission to serve as references, or opt to have CMOs make joint presentations with clients at

conferences, says Marion Schrader, PhD, senior director of marketing at Rentschler. Despite what each company decides to

do regarding information sharing, Schrader points out that for approved drugs, the name of a CMO that has worked with a

pharmaceutical client can be found in drug-approval documentation.

According to Cynthia Wooge, global strategic marketing at SAFC, companies rarely disclose their supplier relationships

until after the drug is approved and a commercial supply agreement is established. There is a growing trend, however, for

companies to disclose supplier relationships earlier in development, but Wooge says “a significant majority continue to hold

information fast until disclosure is necessary or beneficial later in development.” –Randi Hernandez



can continually test the CDMO/

deve lopment compa ny re la -

tionship,” says Cynthia Wooge,

global st rateg ic market ing at

SAFC. Specifically, the analyti-

cal tasks required for a project

create the greatest chance for

out-of-scope work requests, she

says. “Upfront diligence in defin-

ing the scope, deliverables, and

requirements of a program pro-

vides the best opportunity to

limit out-of-scope occurrences.”

A n out sou rc i ng f i r m c a n

also expand its capabilities by

partnering with a second CMO

for activities such as PEGylation

or antibody conjugation. Cobra

Biologics has such a partnership

with Quiapeg to complement its

service offerings. Other options

inc lude ask ing the c l ient to

perform product-specif ic tasks

it se l f , says Johannes Re ite r,

head of biotech cooperat ions

at S a ndoz , or bu i ld i ng t he

technology at the CMO level,

“but this evaluation is done on

a case-by-case basis only when it

makes sense,” adds Reiter.

According to Gingrich, every

CMO has a list of core capabili-

ties and a running list of out-

sourced service providers they

rely on. “The goal is to match

what can be done in-house and

what needs to be outsourced,”

Gingrich asserts. To establish

a successful partnership with

a client, it’s best for a CMO to

conduct a thorough technical

evaluation of a client request

for proposal (RFP), notes Saffell-

Clemmer, and possibly meet

with the client after review of an

RFP to clarify any outstanding

concerns. Wooge concurs that

managing customer expectations

is crucial: “CDMOs are wise to

apply significant rigor in assem-

bl ing detai led proposals that

clearly articulate the assump-

tions and expected work identi-

fied in the RFP process.”

If a selected CMO cannot per-

form all of the duties required

for the manufacture of a bio-

logic from start to finish, three

to four points of contact with

di f ferent CMOs can be typi-

cal. Smith emphasizes that if a

product were to be developed

from cell-line selection through

to distr ibution to clinical tr i-

als, seven to eight CMO part-

ners could even be probable. For

example, multiple points of con-

tact could conceivably be used

for the following steps: a primary

CMO to handle linker-payload

production, a second CMO to

handle conjugation of the drug

substance, a third to handle the

formulation and aseptic fill/fin-

ish of the drug product, and a

f inal CMO for the purpose of

release and stability testing. This

supply chain design adds a sig-

nificant amount of complexity,

says Alonso-Caplen, more room

for error, and “higher external-

ization risks, especially if [the]

CMOs are global ly situated.”

When it comes to the coordina-

tion of multiple service provid-

ers, Gingrich says that the cGMP

CMO is usually responsible for

the management of all of the

other outsourced activities.

In the c ircumstance that a

customer outsources a drug sub-

stance from a different location

than a drug product, a client

can, and typically will, ask a

CMO to perform a more rigor-

ous API testing and release pro-

gram than it normally would for

a drug substance manufactured

by the customer itself or by a

domestic partner, notes Saffell-

Clemmer.

In the case of biosimilar com-

pa rab i l i t y te s t i ng , mu lt ip le

samples of a drug product are

sourced from various regions or

production sites—and it’s up to

the primary CMO to determine

the degree of similarity between

the originator product and the

biosimilar, notes Gingrich.

1. FDA, Draft Guidance for Industry,

Request for Quality Metrics, (Rockville,

MD, July 2015).

2. The editors of Pharmaceutical

Technology, “A Voice of Their Own,”

Outsourcing Resources, supplement to

Pharm. Technol. 39, pp. s24–30 (2015).

3. MicroProtein Technologies, Inc.,

company homepage, www.mptbiotechs.

com, accessed July 24, 2015.

4. The editors of BioPharm International,

“Biopharma Advances Demand

Specialized Expertise,” Outsourcing

Resources eBook, BioPharm Int., pp.

12–19 (June 2015).

5. BioPlan Associates, Inc., 12th Annual

Report and Survey of Biopharmaceutical

Manufacturing Capacity and Production,

E.S. Langer, Ed. (Apr. 2015).

6. BioPlan Associates, Inc., 11th Annual

Report and Survey of Biopharmaceutical

Manufacturing Capacity and Production,

E.S. Langer, Ed. (Apr. 2014).

7. E.S. Langer, “Fill/Finish Trends,”

Outsourcing Resources eBook,

BioPharm Int., pp. 20–25 (June

2015). ◆




SC

IEP

RO

/Gett

y Im

ag

es

Cell harvesting is a crucial step

in biopharmaceutical manufac-

turing that can have significant

impacts on product quality and

the design of remaining downstream

processes. Growing titers and viable cell

densities, due to improved media and

cell lines, and growing use of perfusion

cell culture are making this downstream

purification step more challenging.

In addition, manufacturers desire to

achieve more efficient, cost-effective pro-

cessing with a smaller footprint. “The

biopharmaceutical market is now fac-

ing increasing demand to be quicker

to market, with lower production costs

and a smaller industrial footprint while

improving productivity,” says Alain

Lamproye, president of the biopharma

business unit of Novasep. New filtration

and separation technologies, including

single-use systems for larger scale har-

vesting operations, are helping manufac-

turers meet these needs.

The move To single useCurrent interest in monoclonal antibody

(mAb) therapeutic candidates produced in

CHO cell expression systems is dominat-

ing development pipelines at major drug

producers, according to Timothy D. Hill,

director of upstream process development

for FujiFilm Diosynth Biotechnologies

USA. “One major shift in mAb production

is the replacement of stainless-steel reac-

tors with single-use bioreactors in order to

streamline operations, decrease change-

over times, eliminate cleaning validation,

and enable rapid capacity expansion as

product demand rises,” he notes.

high Titers and Perfusion Processes Challenge Cell harvesting systems

Cynthia A. Challener

New single-use technologies

and other filtration systems are beginning

to address cost, throughput, and

manufacturing footprint

demands.

Cynthia A. Challener, PhD,

is a contributing editor to

BioPharm International.

Downstream Processing


www.EurofinsLancasterLabs.com

Winner of the CRO Leadership Award for

quality, reliability, productivity and innovation.

Leading experts in:

Chemistry

Biochemistry

Microbiology

Molecular &Cell Biology

Virology

Global Services:

Method Development/Optimization

Validation/Qualification/Transfer

Product Release Testing

Stability Storage & Testing

Raw Materials Testing

Impurities & Residuals Testing

Characterization

Cell Banking

Cell Line Characterization

Viral Clearance

Bioassays

Professional Scientific Services

Perfect timing.

Every time.

The art of mastering cell bank production requires

expertise, innovation and exquisite attention to detail

where every minute counts. With more than 50 years

of combined experience, our passionate, and often

nocturnal, scientists obsess 24/7 to ensure your cell

banks become masterpieces. To that, we offer you:

• Direct access to your scientist for any question, any time.

• Constant oversight and monitoring of your cell banks.

• Continual troubleshooting and status updates during

expansion.

For the most accomplished service in the industry,

trust our cell bank experts to be on your watch.

Cell banking

done right.



In addition to lowering upfront

capita l costs and of fer ing a

higher level of flexibility, single-

use technologies for cell harvest-

ing have also addressed one of the

major issues concerning this pro-

cess step—the risk of cross con-

tamination, according to Frank

Meyeroltmanns, head of product

management for purification tech-

nologies, Sartorius Stedim Biotech.

Single-use technologies also pro-

vide cost-effective drug develop-

ment options for products under

investigation. “Cell removal using

conventional centrifugation drives

up cost due to the high capital out-

lays required for the equipment and

operating costs—which are primar-

ily incurred due to required main-

tenance and cleaning-in-place—are

avoided with single-use technolo-

gies,” Meyeroltmanns adds.

imPaCT of PerfusionContinuous-processing technolo-

gies for both cell culture and down-

stream purification steps are some

of most advanced latest develop-

ments in terms of outcomes, accord-

ing to Lamproye. “In many cases,

continuous processes enable high

productivity while keeping produc-

tion costs low. These types of pro-

cesses also have smaller industrial

footprints and constant productiv-

ity and benefit from on-line control

that automatically adjusts processing

parameters to maintain optimal set-

tings,” he observes.

Improvements in both fed-batch

and perfusion processing have con-

tributed to increasing viable cell

densities and titers and are driv-

ing suppliers to provide improved

clarification technologies so that

recoveries are in line with the

higher outputs of the upstream

process, according to Joe Codamo,

senior project manager for biologics

at Patheon. He adds that continu-

ous processes—both upstream and

downstream—are also attractive as

an approach for reducing unit oper-

ations in order to lower manufac-

turing costs, shorten timelines, and

minimize product losses.

High CHO cell-density cultures

achieved using perfusion bioreac-

tor technology are, in fact, becom-

ing more standard to minimize

the upstream production scale and

reduce the footprint of reactors in the

manufacturing facility, thus lowering

overall capital investment and oper-

ating costs. “Typical peak CHO cell

densities have been reported publi-

cally by manufacturers experienced

with perfusion culture to range from

30–50x106 viable cells/mL culture

medium, with some reports of peak

cell densities over 100x106 viable

cells/mL in experimental bench scale

systems. Furthermore, mAb titers are

reported to exceed 10g/L in produc-

tion cycles of 2–3 weeks,” Hill says.

Thoughts about harvest technol-

ogy have also shifted in response to

adoption of perfusion cell culture

in the industry, according to Hill.

“The goal of harvest has broadened

to include cell and product retention

during production, as well as separa-

tion of product from cells during the

end of production,” he observes.

Challenge of higher TiTersCurrent single-use depth filters need

to handle ever-increasing cell con-

centrations yet deliver continuously

higher yields, but they often suffer

from blockage at lower loading capac-

ities, particularly when such volumes

have high biomass concentrations.

Major process-relevant parameters

for cell cultivation with respect to

clarification are particle-size distribu-

tion and particle quantity, accord-

ing to Meyeroltmanns. “For typical

cell-culture applications, particle sizes

vary between 0.1 µm and 25 µm,

whereas the quantity of total bio-

mass-relevant particles ranges from

10 million/mL to more than 100 mil-

lion/mL. These levels present a chal-

lenge for conventional clarification

techniques, and often multi-step fil-

tration regimes are required that use

up large numbers of filter units and

entail extensive case-based adapta-

tions,” he explains.

Centrifuges are predominantly

used to harvest cells from process vol-

umes above 500 L to reduce cost and

waste, according to Meyeroltmanns.

“Large-scale unit operations up to

2000 L require both scale-up and

scale-down concepts, as well as linear

scalability,” he asserts.

Hill does note that 3M has made

significant improvements to its line

of commercially available tradi-

tional depth filters for cell removal,

and Codamo adds that advances in

depth-filter media have also contrib-

uted to significant improvements

in filter capacity, flux, and product

recoveries. Hill agrees, however, that

depth filtration becomes less cost

effective compared with centrifuga-

tion at CHO cell densities of approxi-

mately 30x106 cells/mL or greater.

new large-sCale, single-use filTraTion TeChnologySartorius Stedim Biotechnologies

has adapted “body-feed filtration,”

(BFF) technology from the blood and

plasma fractionation industry for cell

harvesting to enable large-scale, sin-

gle-use filtration. The new Sartoclear

Dynamics system, which was intro-

duced earlier in 2015, is designed to

replace both centrifuges and depth

filters with a single-step process that

achieves cell clarification of up to

2000-L batch volumes.

“The main principle of BFF tech-

nology involves the addition of a fil-

ter aid. This pharmaceutical-grade

diatomite, a fine diatomaceous earth

(DE) is highly porous, increasing the

permeability of the filter cake that

builds up as clarification progresses

in the filtration system connected

downstream. As a result, the DE pre-

vents filters from becoming blocked,

providing a double advantage of sig-

nificant time savings and high flow

rates,” says Meyeroltmanns. He also

notes that special single-use bags

containing ultrapure DE are also




available for attachment using new,

patented quick connectors and dust-

free DE transfer directly into the cell-

culture fluid in a bioreactor.

“We believe that as a linearly

scalable filtration technology, BFF

closes the biggest gaps by enabling

single-use cell harvesting of fluids

from 2000 L standard single-use bio-

reactors and eliminating the need

for centrifugation technology for

removal of cells from such volumes.

This technology also provides a high

level of flexibility while helping to

significantly reduce upfront capital

investments,” Meyeroltmanns states.

TeChnologies for Cell harvesTing from Perfusion ProCessesPerfusion technology presents the

challenge of retaining cells and prod-

uct within the bioreactor until the

end of the production run, when

the product is separated from the

cells. Hollow-fiber filters initially

developed for the growth of adher-

ent cells have more recently been

used as physical barriers for cell

and product retention, while allow-

ing the passage of fresh and spent

media for exchange in the bioreactor,

according to Hill. He also notes that

single-path tangential flow through

hollow-fiber filters has been replaced

by alternating path flow (ATF, alter-

nating tangential flow) to prevent

cell attachment and filter fouling. By

selecting the appropriate filter pore

size, the process can be highly selec-

tive for operation in the retention or

permeate modes.

The combination of clarification

and product capture into one unit

operation also reduces timelines

and improves product recoveries,

according to Codamo. He points

to Patheon’s expanded-bed adsorp-

tion chromatography technology

as an example.

meeTing fuTure neeDsGoing forward, market needs will

be the main drivers of further cell-

harvesting technology develop-

ment, and particularly the need to

improve the production economics

for biological products, according to

Lamproye. As the biologics indus-

try continues to grow and com-

petition within the industry also

increases with the rise of biosimilars,

Codamo agrees that advances in

cell-harvesting technology will lead

to improved processing efficiencies,

reduced costs, and greater flexibility

for use with all of the mammalian

cell types used in the industry and

the various recombinant proteins

currently in the pipeline.

With respect to the technol-

ogy itself, Meyeroltmanns expects

growing mid-term and long-term

adoption of both large-scale unit

operations and single-use equipment

that provides continuous process-

ing capabilities for CMO environ-

ments and multi-product facilities.

Automation will also be of increasing

importance for biologics production,

according to Hill.

“Particular challenges include the

need to continually improve depth

filter efficiency in single-use applica-

tions to deal with ever-evolving cell

densities and productivities, and the

need to reduce process timelines fur-

ther, especially for products suscepti-

ble to degradation during cell harvest


Harvesting adherent cells for therapeutic applications

Cell-based therapies are still in the early stages of development, and one key

unanswered question relates to harvesting of the cells at commercial scales.

“Harvesting of adherent therapeutic cells is very different from the harvesting of

suspension cells that need to be separated from the desired product. In the former

case, the cell viability and activity of the surface proteins must be retained during

the harvesting process in order to preserve the therapeutic properties of the cells,”

according to Alun J. Fowler, commercial marketing manager for EMEA vaccines

and biologics in the Laboratory Products Group of Thermo Fisher Scientific.

“Currently there is no platform technology available for the harvesting of

therapeutic cells,” he notes. “One factor is the variety of cell-culture techniques

being used; most have very different culture conditions and harvesting

requirements, which makes the development of a platform technology quite

challenging,” says Fowler. Some cells (e.g., stem cells) are grown as sheets

while others are grown on microcarriers, and yet others are intended as

scaffold systems for regenerative medicine. Harvesting of therapeutic cells on

a large scale is also difficult because many of these techniques are not easy to

implement for large quantities of cells, according to Fowler. One possible solution

is massive parallel cell culture as a scale-up approach.

Alternatives to actual harvesting are also being explored, such as detachment

technologies. In one example (the UpCell Surface), adherence to a surface is

switched on and off with changes in temperature, thus allowing the grown cells

to detach as sheets without the need to use enzymes that could affect their

properties. It is possible that harvesting cells as sheets rather than individually

may be cheaper and easier at larger scale.

A second alternative involves the use of biodegradable scaffolds for growth of

therapeutic cells. The plant-based material can be digested by cellulase without

impacting the viability and make-up of the product cells, according to Fowler. He

also notes that enzymatic detachment from microcarriers is being investigated,

but this method requires careful selection of enzymes that will not affect the

crucial aspects of the cell structure.

Contin. on page 39



Single-use technology has been

around for approximately two

decades if disposable capsule

filters are taken into consider-

ation. Aseptic connector devices, mixer

bags, storage bags, and bioreactors fol-

lowed. Currently, single-use systems

(SUS) a lso include disconnectors,

disposable filling needles, and sen-

sors. The technology is still evolving,

and the industry can expect further

advances in the future.

The dynamic nature of SUS provides

excitement for professionals from both

suppliers and end users, who predomi-

nantly believe that SUS is a key technology

for the future of biomanufacturing because

of the following major advantages:

• Flexibility

• Less up-front capital investment

• No batch-to-batch cross contamination

• No re-use cleaning validation

• Less work on scale up

• Shortened time-to-patient.

When end users actually implement

SUS in cGMP biomanufacturing processes,

however, there are still many challenges

including the following:

• Lack of thorough understanding of new

materials (e.g., compatibility with pro-

cessing fluids/extractables)

• Lack of deep knowledge of interaction

between SUS and biologics

• Lack of good practices to prevent leak-

age of SUS (manufacturing/ shipping/

handling)

• Limited interchangeability of compo-

nents (e.g., connectors)

• Limited industrial and regulatory guid-

ance on qualification of SUS.

These challenges have prompted both

suppliers and end users to work collabora-

tively to adequately qualify SUS prior to its

implementation.

SUS ProceSS DeveloPmentSingle-use systems are being used in

almost all stages of biomanufacturing pro-

cesses, including upstream, downstream,

and fill/finish operations. Figure 1 depicts

a typical monoclonal antibody (mAb) drug

manufacturing process. Single-use bioreac-

tors are widely used in cell line expansion,

and the use at production scale is increas-

ing, notably in contract manufacturing.

Single-use buffer filtration system and stor-

age bag application is common in down-

stream processes. Final drug-product sterile

filtration and transfer using SUS has also

being increasingly applied to fill/finish

operations.

The implementation of SUS usu-

ally starts with development of the user

requirement specification (URS). Typically,

the development engineers at biophar-

maceutical companies then work closely

with the design engineers and application

engineers from suppliers to select compo-

nents and standard assemblies as available,

or if not, generate suitable drawings, which

will then be subjected to internal approval

within the end-user company. There are

many factors to be evaluated, including

technical assessment, supply chain, busi-

ness aspects, and supplier quality. PDA

Technical Report No . 66, Application of

Single-Use Systems in Pharmaceutical

Manufacturing, has been developed to

provide comprehensive, high-level guid-

ance about qualification of SUS (1). This

paper focuses on technical assessment in

more detail.

technical conSiDerationS: chemical comPatibilityUnlike traditional stainless-steel equip-

ment, SUS is generally made from poly-

mers including plastics and elastomers.

These polymers are obtained through

best Practices in Qualification of Single-Use Systems

Weibing Ding

The author discusses the current best

practices in technical qualification of single-use

systems.

Weibing Ding, PhD, is principal

scientist Process Development, amgen

inc., thousand oaks, ca 91320

Single-Use Systems


Organised by

CPhI Worldwide hosts over 36,000+ visiting

pharma professionals, 2,500+ exhibitors from

150+ countries and more than 100 free industry

seminars, covering every sector under one roof.

Attending CPhI Worldwide is the most cost

eff ective way to establish new business

relationships, meet with global partners and stay

updated on the latest industry trends.

Mix with the world of pharma, products, people and solutions

Co-located with:

“Being part of the CPhI family to

me is about being part of the very

best the industry has to off er. With

opportunities to meet suppliers,

distributors, logistic specialists and

customers all at one time all in one

place, CPhI off ers us the prospect

to exponentially grow our business.”

Matthew Rosen

CEO Global

Pharmaceuticals

Relief, LLC

Register free and receive your ‘Spain Pharmaceuticals 2015’ Report

13 - 15 October 2015 • IFEMA, Feria de Madrid • Spain

Pre-Connect Congress: 12 October 2015

Join the conversation

@cphiww using #cphiww

l.cphi.com/cphi-worldwide/

Join the world’s leading pharmaceutical event



polymerization of organic mono-

mers in the presence of catalysts. As

a result, they are susceptible to attack

by organic solvents as well as acids

and bases; some, at high concentra-

tions, can even dissolve polymers.

Because biomanufacturing may

involve processing of fluids contain-

ing these chemical compounds, the

chemical compatibility of single-

use components with process fluids

under process conditions should be

the first factor to be evaluated.

Table I lists the common polymers

used in the major single-use com-

ponents. Table II provides the gen-

eral chemical incompatibility of

these polymers with common flu-

ids. Because chemical compatibility

is determined by the concentration

of contacting fluid, the nature of the

polymer, the contact time, and tem-

perature, Table II can only be used

as a general guideline. When one

intends to use SUS with these fluids,

a pre-use qualification test should be

performed to validate that the pro-

cess conditions are not incompatible

with the material in question (2–4).

The consequences of incompatibil-

ity would be leaks from the closed

system (e.g., tubing, connector, chro-

matography column, and biobag),

inaccurate readings from sensors,

failed filtration (filter), and potential

failures in process performance and

product quality.

In addition, pretreatment methods

such as steam sterilization, e-beam

irradiation, gamma irradiation,

vaporized hydrogen peroxide, and

ethylene oxide could have an effect

on the properties of the polymers.

Therefore, the specific process param-

eters of these pre-treatments should

not exceed the supplier’s product

claim, which should be proven by

the supplier’s validation study.

technical conSiDerationS: FitneSS For PUrPoSeSUS used for each unit operation

should be fit for purpose. For exam-

ple, a single-use bioreactor should be

sterile and not adversely affect cell

growth due to certain leachables. It

should provide sufficient mixing and

have acceptable gas exchange, and

should not leak during operation.

For buffer filtration and storage, the

SUS should be sterile, not present any

concerns for extractables/leachables,

and not leak. For the SUS in fill and

finish, it should be sterile, not present

any concerns for extractables/leach-

ables, not leak, and in addition, not

raise any concerns for particulates

and endotoxins.

The questions are: How can we

ensure these requirements? Which

part is the responsibility of the sup-

plier? Which part is the responsibility

of the end user?

In general, process validation

remains the responsibility of the

biopharmaceutical manufacturer.

Supplier data should be appropri-

ately used. Leveraging supplier data

requires the end user to understand

how the supplier data were devel-

oped and assess their suitability to

the processes.

regUlatory reQUirementS For finished pharmaceuticals, FDA

has issued regulations explain-

ing cGMPs in 21 Code of Federal

Regulations (CFR) 210 and 211 (5, 6).

For APIs or drug substances, how-

ever, the agency has not issued reg-

ulations. Nevertheless, the agency

issues guidance documents to articu-

late its current practice on related

topics. Although there is no specific

regulation dedicated to SUS, FDA’s

Q7A Good Manufacturing Practice

Guidance for Active Pharmaceutical

Ingredients (7), which is based on

ICH Q7 (8), is a guidance document

Single-Use Systems

SU component Polymers as materials of construction

FiltersPolyvinylidenefuroride, polyethersulfone, polyamide, polyethylene terephthalate, polypropylene,

polysulfone, polyetherimide

Connectors/disconnectors Polypropylene

Aseptic connectors/disconnectors Polyvinylidenefuroride, polycarbonate, polysulfone, silicone

Biobags Low density polyethylene, ethylene vinyl acetate copolymer

Chromatography columns Polymethylmethacrylate

Tubing Silicone, thermoplastic elastomers

Sensors Silicone, polycarbonate, polysulfone

Filling needles Polyetheretherketone

Table I: Common polymers used in the major single-use components and in contact with process fuids.



that should be studied, understood,

and followed. Components and sys-

tems used for single-use technologies

should be assessed for incoming test-

ing. Key factors listed in the guidance

document (7) include:

“A supplier’s certificate of analysis

can be used in place of performing

other tests, provided that the manu-

facturer has a system in place to eval-

uate suppliers.

“Suppl ier approva l should

include an evaluation that provides

adequate evidence (e.g., past qual-

ity history) that the manufacturer

can consistently provide material

meeting specifications.

“P roce s s i ng a id s , ha z a rd -

ous or highly toxic raw materi-

als, other special materials, or

materials transferred to another

unit within the company’s con-

trol do not need to be tested if

the manufacturer’s certificate of

analysis is obtained, showing that

these raw materials conform to

established specifications. Visual

examination of containers, labels,

and recording of batch numbers

should help in establishing the

identity of these materials. The

lack of on-site testing for these

materials should be justified and

documented.

“Equipment should be constructed

so that surfaces that contact raw

materials, intermediates, or APIs do

not alter the quality of the intermedi-

ates and APIs beyond the official or

other established specifications.”

Failure to follow the guidance

could result in receiving an inspec-

tional observation from regulatory

authorities (9).

SUPPlier PracticeSThe basic document generated by

the supplier and shared with the

end user is the validation guide for

a component or a SUS. At a mini-

mum, the following topics should

be evaluated; for some components,

other specific tests may also need to

be performed:

• Materials of construction (Animal-

derived ingredients free or meeting

the EMA410.01 rev.03)

• Particulates/Endotoxin (United

S t a t e s Pha r macope ia [USP ]

<788>/USP <85>)

• Extractables (USP <661> and

/ or other more robust test

designs [10])

Single-Use Systems

Polymers Incompatible chemical compounds

Polycarbonate High pH fuids, strong acids, chlorinated hydrocarbons, and hot water with constant exposure

Polysulfone Dimethylsulfoxide, dimethylacetamide, chlorinated hydrocarbons, and acetone

PolyethersulfoneDimethylsulfoxide, dimethylacetamide, chlorinated hydrocarbons, acetone, and polyethylene

glycol at high temperature

Polyvinylidenefuroride Dimethylformamide, diethylacetamide, and acetone

Polyamide Low pH fuids

Polyethylene terephthalate High pH fuids

Polyetherimide Methylethylketone and chlorinated hydrocarbons

Low density polyethyleneChlorinated hydrocarbons and some types of detergents/disinfectants that induce stress

cracking

Ethylene vinyl acetate copolymer Concentrated mineral acids, ketones, and chlorinated hydrocarbons

Polymethylmethacrylate Strong acids and alkalis, ketones and chlorinated hydrocarbons

Silicone Concentrated acids and alkalis, methylene chloride, and methylethylketone

Thermoplastic elastomers Chlorinated hydrocarbons

Polyetheretherketone Concentrated mineral acids and halogenated hydrocarbons

*Note: Aromatic and aliphatic hydrocarbons are not considered here because they are rarely used in bioprocessing. The table provides

general guidance, and the actual evaluation should be based on end user’s process conditions

Table II: General chemical incompatibility of these polymers with common fuids in bioprocessing* (see references 2–4).



Single-Use Systems

• Biocompatibility (International

Organization for Standardization

(ISO) 10993-4)

• Cytotoxicity/Bioreactivity (USP

<87>/USP<88>)

• Sterility (USP <71>)

• Transportation validation (ISTA-2A

or American Society for Testing

and Materials [ASTM] D7386-12)

• Shelf life (at least two years post

gamma irradiation)

• Sterilization (ISO11137)

• Integrity or gross leak

• Functional tests.

Functional tests are specific to

each SU component. For bioreactor

bags, film gas barrier tests and film

mechanical strength tests should

Chemical compatibility

Supplier End user

Based on the intended use of the single-use component,

perform chemical compatibility tests and publish the results in

the validation guide. When designing the tests, consider using

worst-case application conditions (e.g., time, temperature, and

pre-sterilization conditions) as well as typical process fuids

(chemical constituents and pH) (12–15).

Evaluate the generic data from supplier. Assess the applicability

of the data to the process. If there is a gap, work out a mitigation

plan, which includes either choosing another component that

is compatible or performing additional testing to confrm the

compatibility within process conditions.

Gross leak

Supplier End user

Perform gross leak test prior to packaging at component

level and system level. Then perform shipping validation

for packaged system after gamma irradiation. For aseptic

connectors, side load testing should be performed to provide

useful information for end user’s actual operation to prevent

potential leakage.

Evaluate supplier approach and data. In some cases, shipping

validation using the actual shipping lane is also warranted.

Perform risk assessment, and perform post-installation pre-use

gross leak test, if deemed necessary based on risk assessment.

During the production, closely monitor if there is leak. After the

production, inspect thoroughly.

Extractables

Supplier End user

Perform extractables study using a recommended standardized

extractables protocol (10) that includes selection of test articles,

test solvents, surface area to volume ratio, pre-sterilization

conditions, test conditions, and analytical methods. Evaluate

effect of extractables on cell growth for bioreactors. When there

is change in materials of construction and/or manufacturing

process, perform extractables assessment.

Evaluate supplier approach and data against the recommended

standardized extractables protocol. Perform a science-based

risk assessment, and perform additional extractables testing,

if there are gaps. Perform toxicity-based safety evaluation as

part of extractables/leachables evaluation. For bioreactors,

perform cell culture growth test, if needed. For drug substance

containers and SUS in fll/fnish, product stability study should

be performed to ensure extractables/leachables do not

negatively interact with drug product.

Particulates and endotoxin

Supplier End user

Perform quarterly testing on master system that includes

representative components. For fll and fnish applications,

perform additional test per lot.

Evaluate supplier approach and data. Perform science-based

risk assessment, and perform additional test, if there are gaps.

Change management

Supplier End user

After the launch of SU components with comprehensive

validation guide, when there is any change in materials of

constructions, manufacturing processes and sites, and

test methods, supplier needs to perform science-based

assessment, qualify the change, and provide change notifcation

as well as the qualifcation documentation to end user.

Ensure that a change notifcation agreement is in place with

suppliers (including sub-suppliers) and that such an agreement

provides suffcient time to assess the effects of any changes

on SUS functionality. Evaluate and assess supplier’s change

notifcation and associated change qualifcation documentation.

If the supplier’s assessment is not suffcient, the end user may

have to perform independent testing.

Table III: Best practices in technical qualifcation of single-use systems (SUS).


The CPhI Pharma Awards provide recognition to pharmaceutical companies who turn

inspiration into innovation and celebrate thinkers and creators breaking new ground in

formulation, process development, packaging, technology and more.

The CPhI Pharma Awards return with eight categories in 2015:

Does your business have a great innovation to

be recognised ?

2015

• Best Innovation in Biologic Drug Development and Manufacturing • Innovation in Supply Chain & Logistic Management • Best Innovation in Manufacturing Technology – in partnership with

Pharmaceutical Technology • CEO of the Year – in partnership with PharmaBoardRoom • Best Innovation in APIs and Excipients • Best Innovation in Packaging • Best Innovation in Process and Formulation Development • Excellence in Partnering & Outsourcing – in-cooperation with

Pharmaceutical Outsourcing Magazine

Advancing Development & ManufacturingPharmTech.com

TM

Submit your entry online by 7th of August and be recognised in the Pharma Industry!

Go to www.cphi.com/awards

SCAN HERESupporting Media Partners



be performed. Carbon dioxide, oxy-

gen, and water vapor transmission

studies can follow ASTM F2476,

ASTM D3985/ASTM F1307, and

ASTM F1249/ISO 15106-3, respec-

tively. Mechanic strength tests

include tensile strength (ASTM

D882/ISO 527), tear strength

(ASTM D1004), seal strength

(ASTM F2097/ISO 15747), brittle-

ness (ASTM D1790), and dynamic

mechanical properties (ASTM

D5026). In addition, drop tests on

bags are performed according to

ASTM D4169-05. For SU assemblies,

the joints between tubing and

barbs should be validated to pre-

vent potential leakage.

In addition to the validation guide,

suppliers may have additional sup-

porting documents, including but

not limited to detailed extractables

report, certificate of quality, and cer-

tificate of gamma doses.

enD USer PracticeSAside f rom qual i fy ing a sup-

plier through a quality audit, a

technical due diligence visit is

necessary to understand the sup-

plier’s manufacturing process,

sources of variation, and their

control. The goal is to ensure

that the supplier can provide SUS

or components with high qual-

ity consistently, which do not

negatively impact the end user’s

biomanufacturing process and/or

drug product quality and safety.

To achieve this goal, technical

transparency is essential. During

the v isit, the technical team

members from both companies

should review test designs, con-

firm test methods/results, review

test repor ts, and understand

the test results. When needed,

sub -suppl iers should a lso be

involved.

The supplier’s release criteria

and end-user’s receiving accep-

tance criteria should be aligned

with each other. These include,

but are not limited to, packag-

ing integrity, appearance/identity,

contamination, and extractables

assessment. The alignment of cri-

teria should include any variabil-

ity in appearance, such as minor

blemishes on the exterior of SUS

arising from transport and han-

dling, which do not impair the

function of the assembly. When

developing specifications, func-

tional specifications should be

taken into account to allow SUS

or component interchangeability

wherever feasible.

The next step is to perform a gap

analysis in the context of the specific

application. The major points to con-

sider are as follows:

• Is the extractables study relevant

and report useful?

• Are the particulate/endotoxin tests

adequate?

• Is the irradiation process validated

and results certified?

• Is cell culture growth in the biore-

actor acceptable?

• Is a gross leak test carried out prior

to irradiation?

• Is shipping validation performed

after irradiation?

• Does the use of the SUS affect

product stability?

Based on the gap analysis, addi-

tional qualification tests/assessments

may be warranted.

beSt PracticeS To ensure successful biomanufac-

turing, the SUS must not leak and

must not adversely affect process

performance and product quality

during production. To ensure the

drug product quality and patient

safety, the SUS shall not be reac-

tive, additive, or absorptive so

as to alter the safety, identity,

strength, quality, or purity of the

drug product (11). The technical

Single-Use Systems

Figure 1: Typical mAb manufacturing process.

DS

DP

Vialthaw

Drug

substance

container

Various

column

purifcation

Inoculum

Formulation Final fll & fnishSterile

fltration

Bioburden

reduction

fltration

storage

UF/DF Protein A

Expansion Harvest RecoveryProduction

Sterilefltration

Viralfltration

ViralInactivation

AL

L F

IGU

RE

S A

RE

CO

UR

TE

SY

OF

TH

E A

UT

HO

R



Single-Use Systems

due diligence should be carried

out prior to the implementation

of SUS. Table III provides a partial

list of best practices to consider

in the technical qualification of

SUS used in biomanufacturing

(12–15).

conclUSionThe partnership between end

user and supplier is essential to

successfully implement SUS in

manufacturing drugs to achieve

success in implementation and

cont inued operat ion, and to

ensure the quality and safety of

the drug products. The supplier

needs to be fully transparent

about materials of construction,

supply chain, manufactur ing

process, sources and control of

variability, impact of changes,

and qual i f icat ion/va l idat ion.

The end user needs to perform

quality audits and technical due

diligence visits to ensure com-

pliance as well as a deep under-

standing of all these aspects, and

then carry out science-based risk

assessment considering the prox-

imity of SUS to the final drug

product container closure system

by applying a phase-appropriate

approach. Any gaps found should

be bridged or filled in to ensure

the successful implementation of

SUS in biomanufacturing.

acknowleDgmentSThe author greatly appreciates

Duncan Low’s, Executive Director,

Process Development, Amgen,

guidance and constructive review

of the manuscript. In addition,

the author sincerely thanks the

collaboration from Amgen’s Raw

Materials & Devices, Supplier

Qual ity Management, Global

Strategic Sourcing, and Process

Development, as well as major sup-

pliers for SUS.

reFerenceS1. PDA, Technical Report No.66, Application

of Single-Use Systems in Pharmaceutical

Manufacturing (PDA, 2014).

2. H. Domininghaus, Plastics for Engineers,

Materials, Preperties, Applications

(Hanser Publishers, 1993).

3. Plastics Design Library, Chemical

Resistance, Second Edition (William

Andrew Inc., April 1996).

4. P. A. Schweitzer, Corrosion Resistance

Tables, Third Edition, Revised and

Expanded (Marcel Dekker, Inc., 1991).

5. FDA, Part 210 Current Good

Manufacturing Practice in Manufacturing,

Processing, Packing, or Holding of

Drugs; General, www.accessdata.

fda.gov/scripts/cdrh/cfdocs/cfcfr/

CFRSearch.cfm?CFRPart=210

6. FDA, Part 211 Current Good

Manufacturing Practice for Finished

Pharmaceuticals, www.accessdata.


cfrsearch.cfm?cfrpart=211

7. FDA, Guidance for Industry, Q7A Good

Manufacturing Practice Guidance for

Active Pharmaceutical Ingredients,

www.fda.gov/downloads/Drugs/

uidanceComplianceRegulatoryInformation/

Guidances/UCM073497.pdf

8. ICH, Harmonised Tripartite Guideline, Q7

Good Manufacturing Practice Guide for

Active Pharmaceutical Ingredients, www.

ich.org/fileadmin/Public_Web_Site/

ICH_Products/Guidelines/Quality/

Q7/Step4/Q7_Guideline.pdf

9. FDA, Form 483, 2013, www.fda.gov

/downloads/AboutFDA/CentersOffices/

fficeofGlobalRegulatoryOperationsand

Policy/ORA/ORAElectronicReadingRoom/

UCM377864.pdf

10. W. Ding et al., Pharmaceutical Engineering

(November/December 2014), pp74-85

11. FDA, CGMP for Finished Pharmaceuticals;

21 CFR 211.65(a), www.accessdata.


CFRSearch.cfm?fr=211.65

12. V. Vinci and S Parekh, Handbook

of Industrial Cell Culture:

Mammalian, Microbial and Plant

Cells (Humana Press, 2003).

13. P. Culter, Protein Purification Protocols,

Second Edition (Humana Press, 2004).

14. F. Jameel and S. Hershenson,

Formulation and Process Development

Strategies for Manufacturing

Biopharmaceuticals (Wiley, 2010).

15. L. Ducry, Antibody-Drug Conjugates

(Humana Press, 2013). ◆

and in the presence of host-cell

impurities,” Codamo observes. He

does note, however, that these issues

have been addressed to some extent

by newer technologies, but further

improvements will be needed. One

example is the development of sin-

gle-use technologies that are better

suited for low-temperature harvesting

and thus would support improved

product quality.

For cell harvesting with perfu-

sion processes, Hill believes that

while ATFs currently provide a good

level of automation, the equipment

is still cumbersome to setup, ster-

ilize, and operate. Cell fouling of

hollow-fiber filters has been the

most frequent problem at large scale

(2000 L). “The concept of separation

of product from cells in a continuous

or semi-continuous manner is neces-

sary on the front end of manufac-

turing to provide a constant process

flow through purification,” he also

notes. Presterilized single-use filter

units would also greatly improve

robustness and build efficiency for

setup and operation of ATF systems,

according to Hill.

He adds that technologies are

also needed to increase the speed

of product separation from cells

through hollow f iber f i lters,

because currently this process is

much slower than continuous cen-

trifugation equipment (up to 24 hrs

for ATF compared with less than 2

hours for continuous centrifuga-

tion at the 2000-L scale). Repligen

Corp., which manufactures ATF

systems, expects to launch a single-

use hollow fiber filter within the

next 12 months, according to Hill,

but he is not currently aware of

any technology under development

that would improve the harvest

time of ATF systems. ◆

Downstream Processing—Contin. from page 31



Biophysical binding studies

ut i l i z ing sur face plasmon

resonance, biolayer interfer-

ometry, isothermal titration

calorimetry, or related techniques are

central to the selection and optimi-

zation of biotherapeutic candidates

based on proteins, immunoglobulin

G (IgG), or advanced formats, includ-

ing bispecifics and fusion proteins.

Though these screening and char-

acter ization technologies are well

established, in a variety of circum-

stances, binding measurements may

be ambiguous or even fail to provide

useful data. This article discusses a

suite of complementary techniques,

all based on light scattering, that are

useful in troubleshooting many of the

underlying characterization issues.

These techniques can help investiga-

tors assess solution quality prior to

running binding experiments, qual-

ify aggregation behavior of analytes,

and character ize complex interac-

tions that may not be amenable to

standard characterization method-

ology. Judicious use of a biophysical

light-scattering toolkit is essential for

robust and reliable interaction studies.

Common problems in interaCtion measurementsExperienced practitioners of biophysi-

cal techniquesÑdesigned to measure

biomolecular interactions, such as

surface plasmon resonance (SPR), bio-

layer interferometry (BLI), isothermal

titration calorimetry (ITC), and oth-

ersÑwill recognize the following not-

uncommon scenarios:

• Poor fits of the signals to theoretical

binding curves

• Ambiguous determination of the

appropriate association model

• Erratic or irreproducible data

• Results that vary with immobiliza-

tion chemistry or chip coating

• Resu lt s that depend on which

binding partner is immobilized or

titrated

• Results that vary greatly from lot to

lot of reagent

• Fouling of microfluidic channels.

The causes of these dif f icult ies

are var ied, so it is convenient to

divide the most common into two

categor ie s : qua l i t y cont rol a nd

method limitations.

Quality control issues typical ly

arise from suboptimal sample prepa-

ration, purification, or formulation.

Manifestations may include aggre-

gates, particulates, other impurities,

concentration-dependent self-associ-

ation, or undesirable analyte surface

adhesion (e.g., stickiness). While sam-

ple quality control problems tend to

be more prevalent during early phases

of expression, purification, and pro-

cess development of a biomolecule,

established processes and commer-

cially available reagents are by no

means immune to such excursions

from the normal standard of quality.

M e t h o d l i m i t a t i o n s i n c l u d e

those imposed by su r face -based

methodologies and those imposed by

indirect interaction reporter signals.

The former are generally related to

l igand immobil izat ion, while the

latter is common to many types of

solut ion-based techniques. Every

experimental technique is subject

to specific pros and cons, and often,

the application of several techniques

is necessary to complete the picture

of an interaction or to cross-validate

results. This is particularly true when

the complexes that form go beyond

standard homo/heterodimer or trimer

stoichiometries.

enhancing protein binding studies with a light-scattering toolkit

Daniel Some

Light-scattering techniques

are useful for interaction

studies when traditional

methods of analysis do not suffice.

Daniel Some, PhD, is principal scientist

and director of marketing at Wyatt

technology Corp., [email protected].

protein Characterization



AL

L F

IGU

RE

S A

RE

CO

UR

TE

SY

OF

TH

E A

UT

HO

R.

light sCattering: the solution for CharaCterizing solutionsMa ny of t hese t rouble some

phenomena may be avoided

or overcome by applying one

or more characterization tech-

niques based on light scatter-

ing. Described in the following

examples are the light scatter-

ing tools most complementary

to biophysical interaction analy-

sis: size-exclusion chromatogra-

phy–multi-angle light scattering

(SEC–MALS), dynamic light scat-

tering (DLS), composition-gradi-

ent–multi-angle light scattering

(CG–MALS), and composition-

gradient– dynamic l ight scat-

tering (CG–DLS). Each of these

tools consists of a sample prep-

arat ion and del ivery method

combined with one of the two

primary flavors of light scatter-

ing for solution character iza-

tion: multi-angle light scattering

(MALS) and dynamic light scat-

tering (DLS).

malsMALS is a first-principles tech-

n ique fo r de te r m i n i ng t he

molecular weight and size of

macromolecules and nanopar-

ticles in solution/suspension (1).

A beam of light impinges on the

solution. While most of the light

traverses the solution volume

unimpeded, a f ract ion of the

photons meet solute molecules

and scatter in a l l d irect ions.

The magnitude R of scattered

intensity relative to the incident

intensity (excess Rayleigh ratio)

is directly related to the molecu-

lar weight of the solute M, the

solute’s mass/volume concentra-

tion c, and the relative differ-

ence in refractive index of the

solute and solvent dn/dc. There

is also an angular dependence to

the scattered intensity when the

particles are larger than approxi-

mately 25 nm in diameter, but

because most proteins and other

biomolecules involved in inter-

act ion studies do not exceed

this size limit, we will hereafter

ignore the angular dependence.

The fundamental thermody-

namic relation, which may be

considered the “ideal gas law”

for light scattering—representing

the limit of a dilute system of

point-like particles—is expressed

as R=K·M·c∙(dn ⁄dc)2 where K is a

system constant. By measuring

the scattered intensity and the

concentration, MALS provides

a first-principles determination

of molar mass. When more than

one species of solute is present,

MALS measures the weight-aver-

age molar mass Mw=∑iMici/∑ici .

DlsDLS is a f irst-pr inciples tech-

n i q u e f o r m e a s u r i n g t h e

diffusion coefficient of macro-

molecules and nanopar t ic les

in solution/suspension and for

est imat ing par t ic le sizes and

size distr ibutions (2). As in a

MALS measurement, in DLS, a

laser beam impinges on the solu-

tion and scatters from particles.

Unlike in MALS, a DLS measure-

ment does not consist of simply

measuring the average scattered

intensity, but in determining its

rate of f luctuation. These f luc-

tuations arise from the scatter-

ers’ Brownian motion, the rapid

jittering of particles buffeted by

solvent molecules. Because the

rate of fluctuation is correlated

to the particle’s diffusion rate,

diffusion coefficient(s) may be

determined by analyzing the

DLS fluctuations. A measure of

particle size known as the hydro-

dynamic radius rh is calculated

from the Stokes–Einstein equa-

tion rh=(kBT ) ⁄(6πηDt).

When more than one popula-

tion of particle size is present

in the solut ion, DLS analysis


Figure 1: Size-exclusion chromatogram of bovine serum albumin (BSA) and

hemoglobin (Hb). Points indicate molar masses measured by on-line multi-angle

light scattering (MALS) (DAWN, Wyatt Technology) at each elution time. The

identifcation of the early peak as BSA dimer is confrmed by its molar mass

relative to the monomer. The late elution time of Hb would erroneously indicate a

much lower molecular weight than BSA. This pitfall is avoided by use of MALS,

which also indicates the decreasing molar mass across the Hb peak’s trailing

shoulder; this is a result of dynamic dimer-tetramer dissociation in the Hb complex.

200,000

160,000

120,000

80,000

40,000

32.0 36.0 40.0 44.0

Mo

lar

ma

ss (

g/m

ol)

Time (min)

BSA monomer

(66 kDa)

BSA dimer

(133 kDa)

Hemoglobin

(52-62 kDa)



may ref lect their distribution.

Depending on the range of sizes,

the d ist r ibut ion i s t y pica l ly

described in one of two ways:

• Polydispersity: If the size range

is l imited around a central

value, DLS reports the aver-

age size and the polydispersity

index, which represents the

width of the distribution.

• Multi-modal distributions: If

size populations are separated

by a factor of 3–5x, a distribu-

tion exhibiting distinct peaks

around each size range may be

calculated. The polydispersity

index of each peak may also be

assessed.

SEC –MALS uses SEC coupled

w ith MALS to determine an

accurate distribution of solution

molecular weights of a sample

from first principles. SEC sep-

arates molecules according to

hydrodynamic size, and in stan-

dard SEC analysis, the reten-

tion time is related to molecular

weight via a series of calibration

standards. Hydrodynamic size

may or may not be re lated

directly to molecular weight; the

specific size/weight relationship

depends on a molecule’s con-

formation. Additionally, mole-

cules may interact non-ideally

with column-packing materials,

or columns may age, leading to

skewed retention times. Hence

standard SEC calibration using

globular, hydrophilic proteins

often does not ref lect the true

molecular weight of the elut-

ing sample, as shown in Figure

1. SEC–MALS overcomes these

limitations by determining inde-

pendently the molar mass of

each elution volume, regardless

of elution time (3). While there

are a variety of important uses

of SEC–MALS for protein charac-

terization, including analysis of

the solution molecular weights

and conjugation state of glyco-

proteins and membrane proteins,

the appl icat ions most useful

in the context of biomolecular

interactions are:

• Determination of a protein’s

oligomeric state in solution

• Assessment of aggregates

• Testing for self-association

• Preliminary analysis of hetero-

complex stoichiometry, espe-

cially when binding affinity is

high enough to survive dilu-

tion in the SEC column.

CG –MALS couples a MALS

detector with a composition gra-

dient (CG) system, which auto-

matically prepares a series of

concentrations or compositions

and injects them sequentially

to the MALS and concentration

detectors, with no accompanying

separation step (4, 5). Each com-

position is analyzed by MALS to

determine molecular weight (Mw),

which increases with the forma-

tion of complexes in a direct and

intuitive fashion with respect

to the nature of the complexes

formed. Hence, dimer formation

results in complexes that scatter

twice as much as the two individ-

ual monomers combined; trimer

formation results in complexes

that scatter three times as much

as the three individual monomers.

A complete CG–MALS analy-

sis determines equilibrium dis-

sociation constants Kd as well as

absolute molecular stoichiome-

tries—the number of molecules

of each type in the complex,

rather than the ratio of moles

of each type that react to form

the complex. Hence, CG–MALS

excels at analyzing self-associa-

tion to determine the oligomer(s)

formed, as well as complicated

interactions involving multiple

complexes, simultaneous self-and

hetero-association, and coopera-

tive interactions—all in solution

without labeling or immobiliza-

tion. As a general rule of thumb,

CG–MALS is appropriate when

the molar mass of the complex

is at least 10% greater than the

largest constituent monomer’s

molar mass.

Batch DLS simply refers to DLS

measurements in a cuvette or

other vessel with no separation.


Figure 2: High-throughput–dynamic light scattering (HT–DLS) analysis

of proteins samples in situ in a 96-well plate (DynaPro Plate Reader, Wyatt

Technology). The analysis is color-coded to highlight different levels of purity,

indicating which solutions will provide high-confdence results and which are not

suitable for further use.

Monodisperse / low aggregation

Polydisperse / moderate aggregation

Signifcant aggregate content


Register for free at

http://www.biopharminternational.com/bp/Purifcation

Monoclonal Antibody Purification by Protein A Affinity and Hydroxyapatite Mixed Mode Multi-Column Continuous Chromatography

LIVE WEBCAST: Wednesday, October 7 2015 at 2:00-3:00pm EDT

EVENT OVERVIEW:

Increasing manufacturing capacity and decreasing cost per

purifcation campaign are crucial factors for making antibody

therapies more afordable. With increased upstream titer comes

the drawback of higher levels of impurities, a burden that can

further stress downstream processes that already account for

up to 70% of monoclonal antibody (mAb) production costs.

Multi-column continuous chromatography (MCC), a form of

simulated moving-bed chromatography (SMBC), is a scalable

technology previously demonstrated to improve productivity

and lower the cost of Protein A afnity chromatography versus

the standard single-column batch process.

This presentation will explain how implementing a continuous

downstream process, through the use of MCC as a cost-reduc-

ing capture step and hydroxyapatite for aggregate removal and

concurrent depletion of impurities following Protein A purifca-

tion of mAbs, can increase efciency and reduce the costs of

mAb production.

Who Should Attend:

n Chromatographers, method developers, and process engineers who are involved with the development of downstream purifcation processes that include Protein A capture and aggregate removal steps.

For questions contact Kristen Moore at [email protected]

Key Learning Objectives:

n Discover how multi-column continuous chromatography difers from traditional chromatographic methods.

n Understand how high-capacity protein A resin and hydroxyapatite, in conjunction with multi-column continuous chromatography, can be an efective means of increasing process efciency and reducing production costs.

n Learn how to select the appropriate conditions to make the most efective use of high-capacity Protein A and hydroxyapatite resins in a multi-column continuous chromatography mAb separation.

Presenter

ANTHONY GRABSKI, Ph.D

Director of Research and

Development

Semba Biosciences Inc.

Moderator

RITA PETERS

Editorial Director

BioPharm International

Sponsored by Presented by




High-throughput–dynamic light

scatter ing (HT–DLS) performs

standard DLS analysis in high

throughput format, in situ, in

standard microwell plates, mak-

ing possible analyses that oth-

erwise would be too onerous to

carry out on a regular basis. Figure

2 illustrates a common solution

quality visualization scheme.

CG –DLS is simi lar in con-

cept to CG–MALS, but the aver-

age size (rather than the average

molar mass, as in CG–MALS) is

measured as a function of con-

centrat ion and/or concentra-

tion to assess interactions (6).

While the range of Kd that may

be quantified via CG–DLS is sev-

eral orders of magnitude less that

CG–MALS, and analysis of sizes

is not as rigorous as analysis of

masses to calculate interaction

parameters, CG–DLS does offer a

few advantages, including much

lower s a mple consu mpt ion,

simple operation in a microw-

ell-plate format, and the ability

to readily carry out temperature

ramps to determine entropy and

enthalpy of the interaction via

van’t Hoff analysis. Perhaps one

of the most useful aspects of

CG–DLS is its ability to quickly

screen hundreds of binding part-

ners, in solution, with a simple

determination or validation of

complex stoichiometry.

Quality Control With light sCatteringClean protein solut ions with

minimal aggregates, particulates,

or other impurities are crucial to

obtaining accurate and repeat-

able binding data. A publication

(7) outlined how poor solution

quality may impact SPR or BLI,

with phenomena that include

noisy signals, unusual/poorly fit-

ting binding curves, low active

concentrations, and even fouling

of microf luidic channels. The

presence of two analytes (e.g.,

monomer and dimer) that bind

to the immobilized ligand, for

example, will lead to a binding

curve that includes two “on”

rates and therefore w i l l not

be well represented by a sin-

gle exponential fit. In another

example, an aggregate contain-

ing multiple binding sites will

often exhibit avidity, or anom-

alously enhanced binding and

slow “off” rates not governed by

the usual fit that assumes a sin-

gle binding site.

Other interaction analysis tech-

niques are no less susceptible to

skewed and erroneous results

caused by poor solution quality.

All solutions (including pure buf-

fer) should be prescreened to test

for aggregation, particulates, etc.

The quickest and easiest means

for assessing solution quality is

batch DLS, which can be per-

formed with as little as 1 µL of

sample (4–20 µL is more typical)

and just a few seconds of mea-

surement time (10–30 seconds

is typical). Batch DLS provides

a low-resolution size distr ibu-

tion covering 0.2–2500 nm in rh,

highlighting the presence of large

particles or aggregates as well as

of low oligomers.

In a qua l it y- d r iven work-

flow, all solutions would be pre-

screened by batch DLS. Solutions

with large particulates would be

filtered or centrifuged to remove

the pa r t ic les , and the solu-

tion rechecked. Buffers should

be cleaned up to see no appre-

ciable particles above ˜0.3 nm.

For protein solutions, if no large

Figure 3: Size-exclusion chromatography–multi-angle light scattering (SEC–

MALS) data indicating dynamic equilibrium between monomers and oligomers, via

the concentration dependence of molar mass (DAWN, Wyatt Technology). Three

injections with different starting concentrations and concentration variation across

each peak all point to rapid, reversible self-association.

140

120

100

80

60

200 μg

cmax = 0.10 mg/mL

100 μgcmax = 0.05 mg/mL

50 μgcmax = 0.02 mg/mL

17 18 19Time (min)

Mo

lar

ma

ss (

kD

a)

20

light-scattering

analyses are

generally

straightforward,

intuitive, and

informative.



particles (i.e., more than 3–5

times the size of the monomer)

remain, then the polydispersity

of the protein peak should be

checked. Because it is possible

to f i lter protein solut ions to

0.02 µm using syringe-tip filters

(Whatman), most larger oligo-

mers may be removed as well.

When large numbers of solu-

t ions a re to be prescreened,

HT–DLS offers a ready solution.

Because it uses standard microw-

ell plates and measures in situ

in the plate with no liquid han-

dling, HT–DLS is ideal for inte-

grating with interaction analysis

technologies that sample from

these plates (SPR, ITC) or make

the measurements directly in the

plates (BLI).

The final quality check of a

protein solution consists of a

SEC–MALS measurement to ver-

ify the oligomeric state of the

protein and determine size and

quant it ies of low ol igomers.

More than one interaction mea-

surement has been led astray by

assuming that the protein was

monomeric in solution because a

single peak appeared in the SEC

run, or because a denaturing gel

assay indicated the monomeric

molecular weight. SEC–MALS

prov ides con f idence i n t he

reagents to be used in the bind-

ing measurement, and the work-

flow should only proceed if an

investigator is satisfied with the

SEC–MALS results.

overComing methoD limitations With light sCatteringSurface-based techniques such as

SPR and BLI necessarily rely on

assumptions regarding the expo-

sure of relevant epitopes to the

solution, which may not hold

in certain circumstances. More

common, though, are confus-

ing phenomena such as avidity

(the anomalously strong bind-

ing to immobilized ligands of

analytes presenting two binding

sites), surface-chemistry-depen-

dent results (of ten related to

the charge of the immobiliza-

tion layer such as dextran), and

complex multi-valent or coop-

erative interactions that cannot

occur when one binding partner

is immobilized.

While the limitations of sur-

face interaction techniques are

genera l ly overcome by solu-

tion-based techniques such as

ITC, f luorescence anisotropy,

or mic rosca le ther mophore -

sis (MST), most of these solu-

tion-based assays are subject to

their own limitations. A com-

mon drawback is the need for

fluorescent labeling, potentially

modifying the interaction. In

addit ion, most solution-based

measurements involve an indi-

rec t repor te r s ig na l t hat i s

assumed to represent the effect

of a binding interaction, but can-

not be unequivocally assigned to

the formation of a specific bio-

molecular complex. For exam-

ple, ITC measures the release or

uptake of heat; while there is a

good probability that this ther-

mal signal is the result of asso-

ciation or dissociation, ITC does

not offer direct proof of com-

plex formation or dissociation

and cannot indicate unambigu-

ously which complex(es) form,

especially when characterizing

self-association. In instances of

purely entropic binding, no ther-

mal signal is available to report

the interaction.

Analy t ica l u lt racent r i f uga -

tion—sedimentation equilibrium

is quite useful for analyzing a

variety of interactions. Its pri-

mary limitation is the long time

required to equilibrate, during

which it is possible that sensitive

proteins degrade.

Use of SEC–MALS, CG–MALS,

and CG –DLS helps overcome


Figure 4: A complete analysis of antibody-antigen self- and hetero-association

by means of composition-gradient–multi-angle light scattering (CG–MALS)

(Calypso, Wyatt Technology). The left and right sections correspond to single-

species, stepwise concentration gradients that determine the molecular weight

and self-association properties of each protein. The center section corresponds

to a stepwise cross-over gradient that gradually reduces the fraction of antibody

and increases the fraction of thrombin, covering the complete range of hetero-

association stoichiometries. The data indicate expected excess scattering over the

signal if no interaction occurs, pointing to increasing molar mass due to complex

formation. The data are analyzed to determine the molecular composition of the

complex and its binding affinity, even if self-association is present.

Antibodygradient

4

3.5

3

2.5

2

1.5

1

0.5

0Lig

ht

sca

tte

rin

g s

ign

al

(kD

a*

g/L

)

0 20 40 60Time (min)

Actual LS signal LS signal if no interaction

80 100 120 140

Thrombingradient

Light scattering

Hetero-association gradient




most of aforementioned l imi-

tations and are excellent com-

plements to other interaction

analysis technologies. In some

cases, CG–MALS may be the only

technique capable of fully teas-

ing out a complex interaction

with relative ease.

For self-association, it is often

simplest to begin with SEC–MALS.

Plots of molecular weight that vary

along the peak in a concentration-

dependent manner and that vary

similarly with the concentration

of the injected aliquot are good

indicators of a self-associating pro-

tein, as shown in Figure 3. Under

certain assumptions, it is even

possible to determine Kd of simple

oligomerization such as dimer or

trimer formation (8). Alternatively,

other techniques may suggest that

self-association occurs. In a robust

approach, the workflow would

continue from an initial self-asso-

ciation indication to a CG–MALS

analysis, which would accurately

determine the oligomers that

form—even if multiple oligomeric

states are involved—and the bind-

ing affinity for each oligomer.

For hetero-associat ion, too,

SEC–MALS may be an excel-

lent initial indicator of complex

stoichiometry. A series of pre-

incubated aliquots prepared at dif-

ferent stoichiometric ratios may be

tested, with analysis consisting of

determining the molar mass of the

eluting complexes and unbound

monomers (2). For example, if the

complex is 1:1, and the aliquot is

prepared at a 1:1 ratio, then little

to no unbound monomer should

remain, but if the complex is

2:1, then there will be an excess

of B and a significant amount of

unbound B should elute in a sepa-

rate peak from the complex. While

surface-based techniques are quite

limited in this respect, other solu-

tion-based techniques also give

good initial indications of hetero-

association stoichiometry.

A robust workf low designed

for fully characterizing a het-

ero-association should proceed

to a CG–MALS analysis such as

that of Figure 4. CG–MALS will

validate the true molecular stoi-

chiometry of the complex and

determine Kd by fitting the data

of Mw vs. composition under the

correct association model.

A variety of advanced interac-

tions are not addressed well by

most techniques. These include

simultaneous self- and hetero-

association, interactions between

t wo mult i -va lent molecu les ,

higher-order self-assembly, and

cooperative effects that lead to

formation of higher-order com-

plexes. Because CG –MALS is

a f i r s t-pr i nc iple s , solut ion-

based technique that provides a

direct reporter signal (molecular

weight), it is well suited for tack-

ling such challenging interactions

and providing a comprehensive

analysis. In parallel, SEC–MALS

may provide an initial indica-

tion of what forms are present in

solution, guiding the CG–MALS

method design.

CG–DLS may be used in a simi-

lar manner as CG–MALS, with a

lower range of measurement and

less robust analysis albeit much

lower sample consumption. CG–

DLS becomes limited in terms of

analysis when the order of the

interaction becomes high, since the

relationship between stoichiometry

and rh may become ambiguous.

summaryInteractions between molecules are

complicated. Careful characteriza-

tion of reagents, as well as a criti-

cal examination of binding studies,

is necessary to guarantee accurate

and repeatable results, regardless of

the techniques applied. Many of the

challenges related to both reagent

characterization and validation

of interaction measurements may

be addressed successfully through

a suite of analytical tools involv-

ing light scattering. Light-scattering

analyses are generally straightfor-

ward, intuitive, and informative.

They solve analytical questions with

minimal time and effort, enhance

productivity, and minimize ambi-

guities in interaction studies.

referenCes 1. P.J. Wyatt, Analytica Chimica Acta

272, pp. 1–40 (1993). 2. I. Teraoka, Polymer Solutions: An

Introduction to Physical Properties, (John Wiley & Sons, Inc. ISBN 0-471-38929-3, New York, NY, 2002).

3. J. Wen, T. Arakawa, and J.S. Philo, Analytical Biochemistry 240, pp. 155–166 (1996).

4. D. Some and S. Kenrick, “Characterization of Protein-Protein Interactions via Static and Dynamic Light Scattering” in Protein

Interactions, J. Cai and R. Wang Eds. (InTech, 2012), pp. 401–426.

5. D. Some, Biophysical Reviews 5 (2), pp. 147–158 (2013).

6. A. D. Hanlon, M. I. Larkin, and R. M. Reddick, Biophysical Journal 98 (2), pp. 297–304 (2010).

7. D. Some, “Protein Quality Control in SPR and BLI High-Throughput Screening Studies,” www.wyatt.com/files/literature/white-papers/protein-quality-control-high-throughput-screening-studies.pdf, accessed Aug. 14, 2015.

8. S. Das et al., J. Bacteriol. 190, pp. 7302–7307 (2008). ◆

Clean protein

solutions with

minimal aggregates,

particulates, or

other impurities are

crucial to obtaining

accurate and

repeatable binding

data.


eve

ryth

ingpossib

le/G

etty Im

ages

SPECIAL SESSION

CPhI Pharma Insights Briefing

The Outlook for the Bio/Pharmaceutical Contract Manufacturing Industry

Presented by Jim Miller, President, PharmSource

Wednesday, October 14 from 12:00 – 12:45 PM

While contract manufacturing organizations (CMOs) consolidate their positions in traditional markets and

technologies, they are not addressing some of the most important trends in pharmaceutical manufacturing.

Learn about these trends, as well as the status of the CMO industry, opportunities it offers to clients, and the

challenges it must face to remain relevant in the future.

Located in The Pharma Forum, outside of Halls 5 & 6.

GEt tHE LAtESt

COVERAGE OF:• ProcessDevelopment

• ManufacturingTrends

• Formulation

• AnalyticalTechnology

• RegulatoryCompliance

• QualityAssurance

• APIsynthesis

• Packaging

• Outsourcing

• IndustryBestPractices

PHARMACEUtICAL

tECHNOLOGY EUROPEoffersprint&digital

subscriptions

FREE PRINt & DIGItAL

SUBSCRIPtIONS

www.pharmtech.com/

subscribe-pharmtech

www.PharmTech.com/

LinkedIn

www.twitter.com/ PharmTechGroup

SPECIAL EDUCATIONAL PROGRAMMING

PHARMACEUTICAL TECHNOLOGY EUROPE IS tHE

LEADING PHARMACEUtICAL DEVELOPMENt &

MANUFACtURING PUBLICAtION IN EUROPE.

WE HAVE BEEN PROVIDING OBJECtIVE AND

RELIABLE EXPERt EDItORIAL COVERAGE IN tHE

BIO/PHARMACEUtICAL INDUStRY SINCE 1989!

Come See US at Booth 1a3

SIGN UP FOR YOUR FREE SUBSCRIPTION TODAY!

Organised by:


INTERNATIONAL

Come see

these UBM

publications

at Booth 1A3



So

rend

ls/G

ett

y Im

ag

es

To maintain a state of control

and comply with regulatory

authorities, many pharma-

ceutical, biotech, and med-

ical-device companies have adopted

continued process verification (CPV)

initiatives for manufacturing pro-

cesses (1). By adopting a proactive

approach to monitoring, life-science

manufacturing companies can iden-

tify changes in their manufacturing

processing prior to a quality event

such as a batch failure. Control charts

are the predominant monitoring tool

used to monitor parameter data across

batches with control limits defined

for the normal manufacturing pro-

cess. Control chart run rules generate

signals when non-random processes

occur and flag potential process issues.

The maximum partial pressure of

CO2, for example, may have an unde-

sired upward trend due to a machine

calibration issue, and a signal would

prompt a process engineer to investi-

gate before drug potency was affected.

Valid signals can reduce costs, improve

process understanding, and enhance

operational reliability (2).

Despite the well-defined benefits of

run rules, many life-science companies

misuse them due to lack of guidelines

and/or statistical expertise regarding

statistical process control, which may

lead to incorrect signals (i.e., the inabil-

ity to differentiate between signals and

random noise) and, ultimately, failure

of the monitoring system. Essentially,

the goal when using run rules is to

ensure that valid signals are generated,

Utilizing Run Rules for Effective Monitoring in Manufacturing

Aaron Spence

To enable efficient

monitoring systems, life-

science companies

need to effectively

apply run rules.

Aaron Spence is manager of

analytic consulting, BIOVIA.

Process Controls



correct signals are not overlooked,

and false signals are not created.

This article discusses the life-

science manufacturing industry’s

current use of monitoring tech-

niques and provides guidance

on how to improve the value

obtained from monitoring pro-

grams and run-rule signals.

CURREnt IndUstRy PRACtICEsLife-science manufacturing compa-

nies with mature CPV initiatives

apply run rules to critical/key pro-

cess parameters and quality attri-

butes for trending purposes (4).

Despite small variation among

companies, run rules are typically

applied to parameters based on

risk assessments (e.g., process fail-

ure mode and effects analysis, esti-

mated process capabilities, etc.).

Two common issues life-science

companies often face in the early

stages of adopting a CPV initia-

tive are over-alerting (e.g., generat-

ing false, non-value-added signals)

or under-alerting (e.g., overlook-

ing valid signals). Over-alerting

is often the result of monitoring

every parameter with every run

rule. Conversely, under-alerting

can occur when companies only

evaluate if critical release param-

eters fall within specification lim-

its and adequate monitoring is not

performed. The objective of run

rules in a mature CPV program is

to generate valid signals that pro-

vide useful information for engi-

neers and process experts.

One of the principal benefits

companies receive from using run

rules is the enablement of a mon-

itor ing-by-exception solution.

Instead of manually reviewing

control charts on a regular basis,

selected software programs can

automatically alert users to run-

rule signals (i.e., when a run rule

is violated). Then, resources may

be allocated appropriately to eval-

uate parameters with signal viola-

tions. A monitoring-by-exception

solution that can scale up to mul-

tiple sites and products is essential

to a CPV model.

Regardless of their specific prac-

tices, companies should clearly

document the proper use of run

rules in their monitoring proce-

dures to avoid significant regulatory

risk, and responsible individuals

should be assigned to create and

maintain these crucial documents.

Implementation will also likely

require ongoing evaluations and

support from the company.

GEttInG thE RIGht sIGnAlsGenerating valid run-rule signals

provides a variety of benefits,

primarily through cost reduc-

t ions and deployment of an

early-warning system to prevent

product quality issues. Obtaining

valid signals to drive these busi-

ness benefits, however, is depen-

de nt on f ive mat he mat ic a l

assumptions: parameters must

be baselined from an in-control

process, baselined on an accept-

able sample size, detailed regard-

ing data prec ision, normal ly

distributed, and independent.

Receiving correct value-added

signals requires the use of his-

torical data to establish base-

line control limits for ongoing

monitor ing. Many companies

Process Controls

Table I: Monitoring assumptions checklist.

AssumptionAssessed?

Typical assessments Suggested correction(s)

In-control process Process knowledge, outlier identification Remove special cause for baseline

Sample size ≥ 25 or 30 batches availableObtain additional batches, limit run rules, k-factor corrections (3)

Data precision ≥ 5 different parameter values (excluding outliers)Improve data collection procedures, limit run rules, revise data agreements with contract manufacturing organizations

Normality Shapiro-Wilk, Anderson Darling, Q-Q plotTransformations, non-parametric limits, rational subgrouping

IndependenceDubin-Watson, Ljung Box Q, autocorrelation function/partial autocorrelation function (ACF/PACF) plots

Long-term standard deviation for limits, regression methods (e.g., autoregressive integrated moving average [ARIMA], generalized least squares [GLS]), rational subgrouping



overlook the criticality of base-

lining parameters, which results

in inf lated control l imits and

missed valid signals. To ensure

the process is in control, special

cause variation needs to be iden-

tified and removed during the

baseline process. Special cause

variation results from variability

caused by events outside of the

normal manufacturing process,

such as operator error, power

failure, etc. Eliminating special

cause var iat ion provides l im-

its that more accurately reflect

nominal manufactur ing pro -

cesses. Baselining should occur

after a known process change

occurs (e.g., equipment change,

supplier change, etc.) or on a

regular basis (e.g., annually or

semi-annually) depending on the

availability of staff, the number

of batches produced in a year,

and other parameters. When

one is baselining parameters, at

least 25–30 batches should be

available to generate targets that

accurately reflect the manufac-

turing process. The monitoring

system should allow engineers to

differentiate valid signals from

random process noise, as well.

To produce valid signals that

generate value, an organization

needs to measure and record pro-

cess variability accurately with

sufficient data precision. This is

especially important when the

decimal precision for param-

eters results in a small number

of unique values—an issue that

often plagues the life-sciences

manufac tur ing indust ry. For

example, if pH is reported at a

single decimal precision and val-

ues are always 6.7, 6.8, or 6.9,

run rules cannot be appropri-

ately applied and false signals

may be generated or valid signals

may be overlooked.

For an organization to effec-

tively use control charts and run

rules, parameters need to fol-

low a normal distribution. The

assumption of a normal distribu-

tion relies on the fact that only

1 out of 370 observations will

fall beyond control limits just

by chance (5). Abnormal data

Process Controls

Table II: Common run rules used in the manufacturing industry. Control limits = 3 standard deviations (SD) from mean; warning

limits = 2 SD from mean; inner limits = 1 SD from mean.

Signal Signal typeWestern

electric ruleNelson

ruleFollow-up

1 point outside of control limits

Single event Rule 1 Rule 1Examine other issues or events (e.g., operator error, machine calibration) or any other abnormal observations from other parameters within the batch.

2 out of 3 points outside of warning limits

Shift

Rule 2 Rule 5

Investigate process or supplier changes; utilize group difference tests (e.g., t-tests, analysis of variance models [ANOVAs]) for categorical process variables such as machines, cleanrooms, etc.; conduct correlations with parameter that displays shift.

4 out of 5 points outside of inner limits

Rule 3 Rule 6

9 points on the same side of the central line

Rule 4 Rule 2

6 points in a row increasing/decreasing

Decreasing variability N/A Rule 3Investigate correlations with the parameter that displays the trend; determine if machine maintenance or calibrations are required.

14 points in a row alternating in direction (increasing/decreasing)

Increasing variability N/A Rule 4Investigate correlations with parameter that displays oscillation; conduct group difference tests if multiple populations are represented.

15 points in a row all within the inner limits

In-control process N/A Rule 7Investigate correlations with parameter that displays decreased variability; examine process improvements.

8 points in a row, none of which are within the inner limits

In-control process N/A Rule 8

Investigate seasonality effects or non-random cycles (e.g., changing suppliers, using different machines, etc.); conduct group difference tests; conduct correlations with parameter that displays cycle.



Contin. on page 57

Process Controls

results in an inflated likelihood

that data will fall beyond control

limits (e.g., a one out of 40 chance)

(6). Non-normal distributions are

highly prevalent in life-science

manufacturing processes, and

they are often expected when

processes have lower or upper

bounds (e.g., yield, hold time,

etc.). Not correct ing for this

assumption violation, however,

may lead to false signals, which

have the effect of devaluing the

significance of a violated con-

trol limit. Over-alerting by gen-

erating false signals can result in

employees simply ignoring the

monitoring system.

Non-independent parameters

have narrower control l imits

and are more likely to generate

false signals, resulting in wasted

resources and an inef f ic ient

monitoring system. The assump-

tion of independence assumes

that prev ious batches do not

affect subsequent batches, and

non-independence often indi-

cates there is a pattern in the

data. It is crucial that CPV and

monitoring plans provide direc-

t ions on how to test assump-

tions and correct for assumption

violations so that monitoring-

by-exception solutions generate

appropriate signals.

To achieve the most va lue

from a monitoring system and

generate valid signals, one needs

to continuously evaluate and

correct the assumptions underly-

ing a system. Without these cor-

rections, false signals will waste

company resources and valid sig-

nals may be overlooked. Table I

provides assessments and suggested

actions for the aforementioned

assumptions.

PROPER REsPOnsE tO sIGnAlsLi fe - sc ience companies w ith

mature CPV models derive value

from run-rule signals by imple-

menting appropriate follow-up

practices. When a monitoring

system is configured correctly,

i ncor porat i ng a l l t he s teps

defined in Table I, signals should

be valid and indicate that the

process has changed in some

way. Thus, when a run-rule vio-

lation occurs, it is important to

appropriately follow up on the

signal. Conversely, life-science

manufacturing companies with

immature CPV initiatives spend a

lot of non-value added time chas-

ing false signals, or they revert to

the other extreme of not follow-

ing up on signals at all. Limited

resources and statistical expertise

can make following up on viola-

tions a struggle for many life-

science manufacturers.

It is c ruc ia l to understand

that run-rule signals should be

t reated d i f ferent ly than out-

of-specification quality events.

Quality events require imme-

diate cor rec t ive and preven-

tive actions and investigations,

whereas not every run-rule sig-

Figure 1: Examples of signals denoting process events/changes.

Single event

Nelson rule 1

Cyclical pattern

Nelson rule 2,5,6,8

Oscillation

Nelson rule 4

Increasing variability

Nelson rule 8

Shift

Nelson rule 2,5,6

Trend

Ind

ivid

ua

ls -

Tre

nd

Ind

ivid

ua

ls -

Sh

ift

Ind

ivid

ua

ls -

Sin

gle

eve

nt

Ind

ivid

ua

ls -

Cycl

ica

l p

att

ern

Ind

ivid

ua

ls -

Osc

illa

tio

n

Ind

ivid

ua

ls -

In

cre

asi

ng

va

ria

bil

ity

Batch ID Batch ID Batch ID

Batch ID Batch ID Batch ID

4

3

2

1

0

-1

-2

-3

-4 F8

10

39

F8

10

37

F8

10

35

F8

10

33

F8

10

31

F8

10

29

F8

10

27

F8

10

25

F8

10

23

F8

10

21

F8

10

19

F8

10

17

F8

10

15

F8

10

13

F8

10

11

F8

10

09

F8

10

07

F8

10

05

F8

10

03

F8

10

01

4

3

2

1

0

-1

-2

-3

-4 F8

10

39

F8

10

37

F8

10

35

F8

10

33

F8

10

31

F8

10

29

F8

10

27

F8

10

25

F8

10

23

F8

10

21

F8

10

19

F8

10

17

F8

10

15

F8

10

13

F8

10

11

F8

10

09

F8

10

07

F8

10

05

F8

10

03

F8

10

01

4

3

2

1

0

-1

-2

-3

-4 F8

10

39

F8

10

37

F8

10

35

F8

10

33

F8

10

31

F8

10

29

F8

10

27

F8

10

25

F8

10

23

F8

10

21

F8

10

19

F8

10

17

F8

10

15

F8

10

13

F8

10

11

F8

10

09

F8

10

07

F8

10

05

F8

10

03

F8

10

01

4

3

2

1

0

-1

-2

-3

-4 F8

10

39

F8

10

37

F8

10

35

F8

10

33

F8

10

31

F8

10

29

F8

10

27

F8

10

25

F8

10

23

F8

10

21

F8

10

19

F8

10

17

F8

10

15

F8

10

13

F8

10

11

F8

10

09

F8

10

07

F8

10

05

F8

10

03

F8

10

01

4

3

2

1

0

-1

-2

-3

-4 F8

10

39

F8

10

37

F8

10

35

F8

10

33

F8

10

31

F8

10

29

F8

10

27

F8

10

25

F8

10

23

F8

10

21

F8

10

19

F8

10

17

F8

10

15

F8

10

13

F8

10

11

F8

10

09

F8

10

07

F8

10

05

F8

10

03

F8

10

01

4

3

2

1

0

-1

-2

-3

-4 F8

10

39

F8

10

37

F8

10

35

F8

10

33

F8

10

31

F8

10

29

F8

10

27

F8

10

25

F8

10

23

F8

10

21

F8

10

19

F8

10

17

F8

10

15

F8

10

13

F8

10

11

F8

10

09

F8

10

07

F8

10

05

F8

10

03

F8

10

01

Nelson rule 3

AL

L F

IGU

RE

S A

RE

CO

UR

TE

SY

OF

TH

E A

UT

HO

R



Sve

ta D

em

ido

ff/G

ett

y Im

ag

es

Troubleshooting

Stability testing of biopharmaceuticals

must be performed according to regula-

tory requirements for drug development

programs to establish the re-test/expiry date

for drugs. During such studies, however, it is

not uncommon that atypical or unexpected

data may arise, and given the crucial need

for these studies, effective and rapid response

is necessary to prevent impact on drug-devel-

opment timelines. The cause of any issue

must be identified, and any newly observed

impurities/degradants must be characterized

and assessed for any risk to safety, quality,

and efficacy.

This article presents three common scenarios

observed during stability testing of biologics and

the use of orthogonal analysis, stability experi-

ence, and troubleshooting measures to identify

causes, assess risk, and define subsequent steps in

the development of a biopharmaceutical.

Protein Product and acidic sPeciesIn the first study, Molecule X was a lyophilized

formulation of a protein, a sterile drug product

contained in a vial with an intended long-term

storage condition of +5 °C ± 3 °C. The material

had been tested by gel isoelectric focusing in the

early phase of drug development. The method

that was used was standard and had not been

further reviewed or optimized during the pro-

gram. The isoelectric focusing (IEF) gel profiles

showed unclear banding with poor resolution.

To prov ide g reater understanding of

charged-based variants of the molecule, a

method with greater sensitivity was devel-

oped using imaged capillary isoelectric focus-

ing (cIEF). As expected, cIEF indicated that

the poorly resolved bands on the

IEF gel actually represented four

charged-based variants. These con-

sisted of the main product isoform

that focused at the expected isoelectric point

of the product (i.e., at pI [isoelectric point] 6.8),

two acidic isoforms, and one basic isoform

(Figure 1). This test was validated and incorpo-

rated into the release and stability programs

for the product. During routine testing at the

three-month time point, however, an atypi-

cal isoform was observed that had not been

detected during previous testing procedures or

during method development/validation. The

data indicated that the change in cIEF profile

was accompanied by a change in the ELISA

results for the molecule, thus showing that the

atypical peak impacted product activity.

To further characterize this atypical peak,

the sample was tested against a frozen retained

sample from T=0, by electrospray mass spec-

trometry quadrupole-time-of-flight (ESI MS

Q-ToF) analysis. The data confirmed that the

T=3 sample had an increased level of deamida-

tion of approximately 15% (Figure 2). In addi-

tion, MS/MS sequencing confirmed that the site

of deamidation was located at an asparagine

residue that was located in the complementar-

ity-determining region (CDR) of the molecule.

These data correlated with tertiary structural

changes identified by near-ultraviolet circular

dichroism (near-UV CD), where changes in the

spectral profile were observed. Supporting data

from Fourier transform infrared (FTIR) analysis

showed that secondary structural changes in

the reduction of β-sheets had occurred.

The investigation therefore showed that the

molecule had an amino acid residue that had

an increased propensity to deamidate in the

active domain. Unfortunately, this was not

identified in early development, as the original

method had insufficient resolution to distin-

guish between charged variants.

Based on the results, the product sequence was

reviewed and modified to remove the susceptible

Stability Testing in BiopharmaThree case studies illustrate some analytical methods important for stability testing.

Stella-Christiana Chotou is team

leader of stability services at sGs Life

science services.



residue. The development program

was significantly delayed as various

additional supporting studies were

required. This delay could have been

prevented if an optimized method

had been developed upfront in the

early development phase.

A forced degradation study dur-

ing early development would con-

firm key degradation pathways,

and determine whether such a

method is stability-indicating for

the drug molecule or not. These

degradation data should also be

used in the validation of the cho-

sen method for charge-based vari-

ant analysis of the product.

MonocLonaL igG druG substance and ParticuLates In a second study, Molecule Y was a

recombinant protein drug substance

in a liquid formulation. The drug

product itself was produced follow-

ing the vialing and lyophilization of

the drug substance. The drug sub-

stance had an intended long-term

storage condition of +5 °C ± 3 °C,

and data had been used to establish

a two-year shelf life.

During ongoing and established

stability studies, activity testing

of the drug substance unexpect-

edly failed at T=6 months. This

result did not correlate with his-

torical data or any of the other

data available. During the investi-

gation, it was noted that all testing

for the sample was performed at

site A with the exception of activ-

ity testing, which was performed

on a small aliquot of the sample

at site B. In this case, the shipping

procedure for the sample used in

activity testing differed from the

one normally used. The courier

had intended to ship the sample

to the testing lab directly, as had

been done previously; however,

this time the courier had made an

error and shipped the sample to

an incorrect location. The sample

then had to be transported to the

correct location. The temperature

monitoring data indicated that the

cold chain had been adequately

maintained throughout the ship-

ment. The only item of note was

that because of the need to cor-

rect the delivery of the sample,

the product had been subjected to

additional eight hours of transport

by truck, which had not occurred

in previous shipments.

troubleshooting

Figure 1: Examples of a) an imaged capillary isoelectric focusing (cIEF) electropherogram at T=0 months, and b) of a cIEF

electropherogram at T=3 months.

A0.50

0.45

0.40

0.35

0.30

0.25

0.20

0.15

0.10

0.05

0.00

-0.054.0

Abso

rbance

-4.22

-9.45

4.5 5.0 5.5 6.0 6.5 7.0

pl

7.5 8.0 8.5 9.0 9.5 10.0

Abso

rbance

-4.22

-9.45

pl

0.50

0.45

0.40

0.35

0.30

0.25

0.20

0.15

0.10

0.05

0.00

-0.054.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0

B

AL

L F

IGU

RE

S A

RE

CO

UR

TE

SY

OF

TH

E A

UT

HO

R.

Figure 2: Electrospray mass spectrometry quadrupole-time-of-fight (ESI MS

Q-ToF) data demonstrating deamidation in the complementarity-determining

region (CDR) domain at T=3 months.

T=3

T=0

This species is doubly charged. Therefore, an m/z increase of 0.5 of a doubly charged signal corresponds to a mass increase of 1 Da.

This is consistent with deamidation.

100

100

893

894.03894.75895.64 896.67 897.60

898.15899.91

900.43

900.92

901.43

901.93

900.43

900.94

901.43

901.93

910.93

902.42

902.62 904.71904.98 906.03907.93

908.41909.44

909.90 911.91

894 895 896 897 898 899 900 901 902 904903 905 906 907 908 909 910 911

893 894 895 896 897 898 899 900 901 902 903 904 905 906 907 908 909 910 911m/z

m/z0

0

%%



troubleshooting

Based on this, a small-scale agi-

tation study was performed in

which the product was subjected

to agitation at 30 rpm over 0, 4,

8, 16 and 24 hours. Samples were

tested using low-volume methods

to assess levels of visible particles

by visual appearance, sub-visible

particles by light obscuration and

smaller size particles by size exclu-

sion chromatography (SEC) and

dynamic light scattering (DLS).

DLS was used to assess the lev-

els of higher-molecular-weight

(HMW) species at particle sizes

from 10nm to 1μm. The results

showed significant levels of higher

MW species in the later time

points compared with the con-

trol sample at T=0 hours, which

was demonstrated in a series of

correlograms (Figure 3). The con-

trol sample showed a steep sig-

moidal correlogram as a result of

the correlogram function, indi-

cating small monomeric-type spe-

cies, whereas the degraded sample

showed a broad, slow decay, indi-

cating the formation of multiple,

large, slowly diffusing, high-MW

species. It was also observed in

the sample wells that no parti-

cles were evident in the T=0 hours

well, but it was obvious that there

were particles in the T=16 hours

sample well. These particles were

identified as consisting of dimers,

trimers, and multimers of the

drug molecule, and it was con-

cluded that the material was sensi-

tive to agitation. This meant that

shipment as a liquid was not a

suitable procedure to use for this

drug product.

Following confirmation that the

product was stable after a freeze/

thaw test procedure, the ship-

ment conditions for the material

were selected as frozen. This issue

could have been identified during

early development had an agita-

tion assessment been performed. It

is recommended that an agitation

assessment be performed as part of

an early-phase forced-degradation

study or that it is incorporated into

the first stability study.

HorMone druG Product teMPerature excursionIn a third study, Molecule Z, a hor-

mone drug product in a liquid

formulation, had an intended long-

term storage condition of +5 °C ±

3 °C. The product was in ongoing

Phase II clinical trials, and during

shipment of a batch for use in the

next stage of clinical trials, the tem-

perature that the sample was being

held at dropped to 1 °C, outside of

the required range.

During shipment, customs had

retained the product at the air-

port to review paperwork. Such a

delay is not uncommon; however,

during this period the container

was held outside in cold weather

conditions, and the temperature

in the container dropped to 1 °C

for up to 5 hours (Figure 4). The

cold-chain data for this product

indicated it was not stable upon

freeze/thaw at temperatures rang-

ing from -20 °C to ambient; how-

ever, no data were available at 1 °C.

The regulators were informed and

Figure 3: Comparison of dynamic light scattering (DLS) correlograms at T=0

hours and T=16 hours.

Figure 4: Temperature data during shipment showing the excursion to 1 °C.

1.18

1.16

1.14

1.12

1.10

1.08

1.06

1.04

1.02

1.00

0.98

0.960.10

Inte

nsi

ty A

uto

corr

ect

ion

1.00 10.0 100.0 1.0E+3

Time (μs)

1.0E+4 1.0E+5 1.0E+6 1.0E+7

T=0 T=16

Temp tale data9

8

7

6

5

4

3

2

1

0

Temperature

lower limit

upper limit

Time

Tem

pe

ratu

re (

oC

)

06:0

0:00

07:0

0:00

08:0

0:00

09:0

0:00

10:0

0:00

11:0

0:00

12:0

0:00

13:0

0:00

14:0

0:00

15:0

0:00

16:0

0:00

17:0

0:00

18:0

0:00

19:0

0:00

20:0

0:00

21:0

0:00

22:0

0:00

23:0

0:00



troubleshooting

the clinical trial was put on hold

until the quality of the material

was reviewed.

An urgent excursion study

on the material was performed,

with a representat ive sample

being taken from the batch and

held in a programmed thermal

cycling chamber at +5 °C for 4

hours, followed by 1 °C for 5, 10

and 15 hours. Stability tests were

performed on an unexposed con-

trol sample and on the exposed

samples, with the resulting data

confirming that the product qual-

ity was unaffected following this

level of exposure. The data were

used to provide scientifically sound

justification that product quality

had not been affected and the clini-

cal trials were continued.

Excursion studies for new prod-

ucts are not mandatory. Studies

that include mapping of the

intended shipment routes, how-

ever, can be used to perform

risk analyses and identify any

high-risk excursions in advance.

Conducting a small-scale study to

identify the impact of most likely

shipping events upfront allows

drug developers to be proactive,

rather than reactive, when an

unexpected temperature excur-

sion occurs.

concLusionStability studies are an integral

part of drug development with

str ingent timelines for analyt-

ical testing. However, it is not

uncommon that at y pica l or

unexpected stability data may

arise that cause severe impact

on the drug development pro-

gram. It is crucial, therefore, for

drug developers to take rapid

and effective action that mini-

mizes the risk to their timelines

and satisfies regulatory authori-

ties. As discussed in this article,

it is recommended that potential

risks to the biopharmaceutical

due to temperature excursions

or agitat ion dur ing shipment

are ident i f ied in early devel-

opment rather than late-phase

stages. Furthermore, it is vital

to develop appropriate stability-

indicating analytical methods to

be able to identify degradants

that present potential r isk to

patient safety. ◆

PRODUCT SPOTLIGHT

Biocontainer Assemblies

Provide Repeatable PerformanceMeissner’s FlexGro single-

use biocontainer assemblies,

designed for use with

rocker-style bioreactors,

feature the TepoFlex

polyethylene (PE) multi-

layer film, are available in

up to 50 L and are delivered

presterilized for immediate

use. The TepoFlex film

offers a clean film platform,

devoid of slip agents,

which provides robust

and repeatable performance for cell-cultivation

processes, the company reports. FlexGro

biocontainer assemblies can be customized to

specific end-user requirements via various sampling

and process connection options, or companies

can choose from standard assembly designs.

Meissner Filtration Products

www.meissner.com

Serialization Software Helps

Companies Stay CompliantAdents’ Pharma Suite serialization

software, developed in compliance

with GMP and 21 Code of Federal

Regulations (CFR) Part 11 regulations and

following GAMP5 guidelines, features

track-and-trace capabilities, including

generation of unique barcodes and

importation of serial numbers, affiliations

management for origin identification

(bundle, pallet, etc.), and distribution

of various types of serialized code.

The software is compatible with both information

systems (e.g., enterprise resource planning, manufacturing

execution systems) and existing production and

packaging equipment. Pharma Suite is designed to

address both current and emerging regulations.

The software includes the Adents Pharma Exchange

gateway to transit serialization data, the Adents Pharma

Supervisor to manage and distribute serialized codes,

and the Adents Pilot to direct printing and vision systems

and manage communication with line equipment.

Adents

www.adents.com



New Technology Showcase

Pa

ge h

ea

de

r im

ag

e: A

rth

ur

S. A

ub

ry/G

ett

y I

ma

ge

s

Protein A resin

Protein A resins constitute a substantial cost

in state-of-the-art mAb purification

processes. Factors such as operating cycles,

capacity, and mAb titer can have an impact on

total costs associated with mAb purification.

High capacity TOYOPEARL AF-rProtein A HC-650F resin from Tosoh Bioscience

LLC has a binding capacity of >70 g/L, generating increased product

throughput, reduced operating costs and increased manufacturing

productivity. Tosoh Bioscience, tel: 484.805.1265, chris.manzari@tosoh.

com, www.tosoh.com

BioFlo® 320 From ePPendorF:

the smArter solution

The BioFlo 320 is the next generation in

bioprocess control. Designed as a universal

platform, it combines a new industrial

design, flexibility between

interchangeable autoclavable and single-use vessels, and universal gas

control strategy for both microbial and cell culture applications.

The BioFlo 320 has the right combination of features to get the job done.

Eppendorf AG, tel: +49 40.53801.0, [email protected],

www.eppendorf.com/contact

nuviA cPrime hydroPhoBic

cAtion exchAnge mediA

The Nuvia™ cPrime™

chromatography media are a new

addition to Bio-Rad’s family of

mixed-mode purification products.

The media are designed for process-

scale purification of a wide variety of therapeutic proteins. The

selectivity allows method developers to use hydrophobic and cation

exchange interaction modes to achieve effective purification.

Bio-Rad Laboratories, Inc., www.bio-rad.com/nuvia

celeBrAte PerFormAnce with

the new ex-cell® AdvAnced™ cho

Fed-BAtch PlAtForm By sAFc®

Introducing the next generation in chemically-defined CHO fed-batch media. This contemporary media and feed platform was developed across a wide range of CHO cell lines commonly

used in industrial bio-manufacturing with an emphasis on simple adaptation (regardless of cell bank medium), demonstrated performance with sustained high biomass and maximum titers, and formulations allowing for flexibility to adjust protein quality attributes as needed. For more information or to try a sample, please visit us at www.Sigma-Aldrich.com/CHOperformance, SAFC®

euroFins lAncAster lABs

As a member of Eurofins’ BioPharma

Product Testing Group—the largest

network of harmonized bio/

pharmaceutical GMP product testing

laboratories worldwide—Eurofins

Lancaster Laboratories supports all functional areas of bio/pharmaceutical

manufacturing, including method development, microbiology, process

validation, and quality control throughout all stages of the drug

development process. Eurofins Lancaster Labs, tel. 717.656.2300,

www.EurofinsLancasterLabs.com

sievers 500 rl

online toc AnAlyzer

The Sievers 500 RL Online Total Organic

Carbon (TOC) Analyzer is designed to

monitor the quality of PW and WFI in the

pharmaceutical manufacturing industry.

If you need to monitor organics, choose

the most reliable technology that is also the simplest to install,

validate, operate, and maintain.

GE Analytical Instruments, 6060 Spine Road, Boulder CO 80301,

tel. 303.444.2009, [email protected], www.geinstruments.com/500

0

10

20

30

40

50

60

70

80

2 3.5 5

Residence time (minutes)

DBC of of TOYOPEARL AF-rProtein A HC-650F

Bin

din

g C

ap

ac

ity

for

IgG

(g

/L)

1 g/L

5 g/L

10 g/L

online viABle cell

density monitoring

Hamilton’s Incyte, viable cell density sensor, enables measurement of viable cells without influence from changes in the media, microcarriers, dead cells, or debris. Designed

for use in mammalian cell culture, yeast and bacterial fermentation, its 12 mm diameter, PG13.5 thread and 120 thru 425 mm lengths fit all reactor sizes. Either 2 or 4 sensors connect to the Arc View Controller, which displays, records, and exports measurement data in 4-20 mA, OPC or Modbus formats. Hamilton Company, tel: 800.648.5950, [email protected], www.hamiltoncompany.com/sensors

wuxi APPtec Begins extensive

mAnuFActuring exPAnsion

WuXi AppTec is adding 460,000 sq. ft. of

commercial drug substance and drug product

cGMP manufacturing facilities to its already

extensive CDMO capabilities. The new campus

will house the world’s largest fully disposable

mammalian cell culture production plant (14 X

2,000L fed-batch and 2 X 1,000L perfusion). The facilities will be operational

by Q2 2016. WuXi AppTec, [email protected], www.

wuxiapptec.com/biologics



Process Controls—Contin. from page 51

nal should be investigated. If a

parameter has been relatively

stable with infrequent and ines-

sential signals for example, it is

acceptable to take no immedi-

ate action. However, it is impor-

tant to track parameter signals

over time so that multiple viola-

tions or problematic patterns are

not overlooked. If a signal does

require follow-up, it is crucial

to identify the type of violation

and fol low-up with appropr i-

ate investigative techniques to

detect a root cause. Although a

multitude of process changes can

occur, run rules are designed to

detect some of the more com-

mon s ig na l s : s ing le event s ,

sh i f t s , t r end s , o sc i l l at ions ,

increasing/decreasing variability,

and cycles (see Figure 1).

Identifying the type of signal

and appropriate follow-up tech-

niques will bring significant value

to a business by reducing resource

and time requirements. Each sig-

nal indicates that a different pro-

cess change/event occurred, and

investigations should be tailored

based on the signal (see Table II).

Once a signal has been detected

and a decision has been made to

pursue it, typical follow-up steps

include:

• Confirm the value is correct

and not a data entry error.

• Determine if there is a readily

apparent root cause or if other

issues occurred in the process.

• Discuss the signalwith subject

matter experts.

• Examinethelottraceabilityand

review parameters related to the

parameter of interest.

• Escalatetoothergroupsforfur-

ther statistical investigation.

While run ru les prov ide a

useful tool for detecting some

non-random processes, not all

patterns can always be flagged.

Additionally, signals can be over-

looked for parameters that are

being trended with few run rules.

Thus, it is v ital to review all

parameters regularly (e.g., annu-

ally) to ensure that trends are

not missed. Individuals respon-

sible for conf ig ur ing, main-

taining, and reporting on the

monitor ing system should be

clearly identif ied in company

documents. Properly responding

to signals will result in more effi-

cient use of resources and time.

Additionally, investigating sig-

nals can enhance process under-

standing, which should improve

process monitoring and opera-

tional reliability.

COnClUsIOnProper CPV monitoring is a cru-

cial business value investment

for a company. A monitoring-by-

exception system that generates

valid signals can reduce resource

requirements, proactively iden-

t i f y issues pr ior to a qual ity

event, and create a regulatory

compliant environment. Today’s

life-science industry can improve

productivity and compliance by

developing more mature CPV ini-

tiatives and adopting practices

to ensure that they get the right

signals. Furthermore, performing

appropriate follow-up on valid

signals is vital to an effective and

continuously improving monitor-

ing solution.

REfEREnCEs1. FDA, Guidance for Industry: Process

Validation: General Principles

and Practices (Rockville,

MD, January 2011).

2. BioPhorum Operations Group,

Continued Process Verification:

An Industry Position Paper with

Example Plan, BPOG–Biophorum

Operations Group (2014).

3. R.J. Seely, L. Munyakazi, and

J. Haury, BioPharm Int. 14

(10), pp. 28–34 (2001).

4. J. M. Juran, Juran on Quality by Design:

The New Steps for Planning Quality

into Goods and Services (Simon and

Schuster, New York, NY, 1992).

5. D. Montgomery, Introduction to

Statistical Quality Control (John

Wiley, Hoboken, NJ, 5th ed., 2005).

6. S. Steiner, B. Abraham, and J.

MacKay, Understanding Process

Capability Indices (University of

Waterloo, Ontario, 1997).◆

Process Controls

Ad Index Company Page

BIO RAd lABORAtORIEs Cover tip

EMERGEnt BIOsOlUtIOns 17

ClInIGEn 19

CPhI 33, 37

EPPEndORf nORth AMERICA 11

EMd MIllIPORE 13

EUROfIns lAnCAstER lABORAtORIEs 29

GE AnAlytICAl InstRUMEnts 60

GE hEAlthCARE lIfE sCIEnCEs 5

hAMIltOn CO 9

IntERPhEX 23

sAfC BIOsCIEnCEs sIGMA AldRICh 59

sGs lIfE sCIEnCE sERVICEs 7

tOsOh BIOsCIEnCE 2, 43


EMA Releases Positive Opinion

for GSK’s Malaria VaccineThe Committee for Medicinal Products for Human Use (CHMP) of the European Medicines Agency (EMA) has announced a positive scientific opinion for GlaxoSmithKline’s (GSK’s) malaria candidate vaccine, Mosquirix (RTS,S), for use outside of the European Union (EU) in children aged 6 weeks to 17 months.

While other vaccines tackle viruses or bacteria, RTS,S has been designed to prevent malaria caused by the Plasmodium falciparum parasite, which is prevalent in sub-Saharan Africa (SSA) and also protects against hepatitis B. In 2013, there were an estimated 584,000 deaths from malaria with approximately 90% of these occurring in SSA, and 83% in children under the age of five in SSA, the World Health Organization (WHO) reports.

RTS,S aims to trigger the body’s immune system to defend against the Plasmodium falciparum malaria parasite when it first enters the human host’s bloodstream and/or when the parasite infects liver cells. It is designed to prevent the parasite from infecting, maturing, and multiplying in the liver, after which time the parasite would re-enter the bloodstream and infect red blood cells, leading to disease symptoms, GSK reports in a statement.

Limited effectivenessData from a clinical trial conducted in seven African countries showed that Mosquirix provides modest protection against Plasmodium falciparum malaria in children in the 12 months following vaccination. The vaccine was effective at preventing a first or only clinical malaria episode in 56% of children aged between 5–17 months and in 31% of children aged 6–12 weeks. The efficacy of the vaccine decreased after one year. The safety profile of the vaccine was considered acceptable, EMA reports.

CHMP concluded that despite its limited efficacy, the benefits of Mosquirix outweigh the risks in both age groups studied and the benefits of vaccination may be particularly important among children in high-transmission areas in which mortality is very high.

Because the studies showed that Mosquirix does not offer complete protection, and the protection it provides decreases in the longer term, EMA states that established protective measures, such as insecticide-treated bed nets, should continue to be used in addition to the vaccine.

Next steps in developmentThe vaccine was submitted to EMA under a regulatory procedure (Article 58) that allows EMA to assess the quality, safety, and efficacy of a medicine or vaccine and its benefit-risk balance, although it will not be marketed in the EU. EMA reports that the CHMP worked closely with WHO and regulatory authorities from the relevant countries and applied the same rigorous standards as for medicines to be marketed within the EU.

The EMA approval was a pre-requisite for a WHO policy recommendation for the use of Mosquirix in vaccination

programs. WHO will consider additional factors not addressed by EMA including feasibility of implementation, affordability, cost-effectiveness, and the public health value of the vaccine in relation to other available malaria control measures and vaccines. WHO will provide its recommendations on use of Mosquirix by November 2015. Regulators in the countries where the vaccine will be used will decide on licensing in their jurisdictions.

RTS,S, which was developed in partnership with the PATH Malaria Vaccine Initiative (MVI), is the first candidate vaccine for the prevention of malaria to reach this milestone, GSK reports. RTS,S was created in 1987 by scientists working at GSK laboratories. Early clinical development was done in collaboration with the Walter Reed Army Institute for Research. In January 2001, GSK and PATH, with grant monies from the Bill & Melinda Gates Foundation to PATH, entered into a public-private partnership to develop an RTS,S-based vaccine for infants and young children living in malaria-endemic regions in sub-Saharan Africa.

GSK reports the company has invested more than $365 million to date and expects to invest a further $200 to $250 million until development is completed. Between 2001 and the end of 2014, the MVI, supported by grants from the Bill & Melinda Gates Foundation, invested more than $200 million to advance the RTS,S project.

In a press statement, GSK noted that the company has committed to a not-for-profit price for RTS,S; if approved, the price of RTS,S would cover the cost of manufacturing the vaccine with a return of approximately 5% that will be reinvested in research and development for second-generation malaria vaccines, or vaccines against other neglected tropical diseases.

CMC Biologics to Manufacture

mAbs for the Treatment of MalariaCMC Biologics will manufacture monoclonal antibodies (mAbs) and provide process development services for the PATH Malaria Vaccine Initiative (MVI), CMC announced on Aug. 12, 2015. The company will use its proprietary production cell line for the project, and will test MVI’s investigational mAb for its ability to target circumsporozoite protein (CSP) and protect against infection by Plasmodium falciparum, the parasite that causes malaria.

“We will deliver cGMP material for MVI’s preclinical and Phase I/II clinical studies in a remarkable 12 months—the fastest development timeline in the industry from DNA to delivery,” Gustavo Mahler, PhD, global chief operations officer at CMC Biologics said in a press release.

Based on the success of the project with MVI, the group may hire CMC to investigate additional mAbs that bind to alternate epitopes. “Assuming this initial study is successful, we will evaluate other antibodies targeting novel vaccine targets for their capacity to protect humans from infection and therefore, inform future vaccine development efforts,” confirmed Ashley J. Birkett, PhD, director at MVI.


Vaccines Development Update

Ho

u Y

uxu

an

/Ge

tty I

ma

ge

s


Introducing a next generation, chemically-deÀned CHO fed-batch media platform from SAFC. Developed

across a range of widely used industrial CHO cell lines, this newest portfolio media delivers exceptional

titers and economic efÀciencies. Adaptation is simple. Celebrate performance and accelerate your

bio-development process.

Celebrate! Request your complimentary sample at

sigma-aldrich.com/CHOperformance

Cat. Nos: 14366C, 24366C, 24367C, 24368C

EX-CELL® ADVANCED™ CHO FED-BATCH SYSTEM

CELL CULTURE

PERFORMANCE

©2015 Sigma-Aldrich Co. LLC. All rights reserved. Sigma-Aldrich, SAFC, and EX-CELL are trademarks of Sigma-Aldrich Co. LLC, registered in the US and other countries.

NEXT GENERATION MEDIA PLATFORM


www.geinstruments.com/M9

Your workday can be a chaotic place, tools that boost

productivity are essential. That’s why Sievers M9

Total Organic Carbon (TOC) Analyzers are the industry

standard—created in Colorado, USA by people who

know a little something about the best way to tackle

a daunting climb.

Time-tested dependability, now twice as fast

Always quick to set up and easy to use, the Sievers

0��/DE��2Q�/LQH��DQG�3RUWDEOH�$QDO\]HUV�RȖHU�WZLFH�

as-fast readings, smarter data management, easy

maintenance, and an instinctively simple interface.

Enjoy peace of mind using the perfect tool for

PRQLWRULQJ�SXUL¿HG�ZDWHU��ZDWHU�IRU�LQMHFWLRQ��DQG�

cleaning validation samples. Simplify your organics

testing and improve productivity with an M-Series

TOC Analyzers.

.

Sievers M9 TOC Analyzers Reach the peak of productivity

GE Power & WaterWater & Process Technologies

Analytical Instruments

Now with Sample Conductivity!

To view a short video of the Sievers M-Series TOC

Analyzers, scan the QR code or go to the website below.


cmo specialize to build better biologic...

Documents