robert morenweiser - dechema · the world vaccine market 93% 7% pharma vaccines 53% 47% 2006 data...

Vaccine ManufacturingChallenges & Strategies

Robert Morenweiser

Fast Trak Services Europe

GE Healthcare

2Dechema Vaccine Workshop_Oct 2010

© 2010 General Electric Company – All rights reserved

Outline

• Challenges in Vaccine Production

• Current Trends & Correspondings Strategies

• Summary



The world vaccine market

93%

7%

Pharma Vaccines

53%

47%

2006 Data

Pharma market: 241 Bn $

Vaccine market: 18 Bn $

89%

11%

non USA USA

Industrialized countries

Developing countries

Ref. Human Vaccines 2007 Reportwww.strategyR.com



Why this enormous interest in Vaccines?

Vaccine environment

No 1 solutionto global health issues

New technologies

Greater knowledge ofDiseases & immune system

New emerging markets

Need to cut Healthcare spending

Pandemic threat /Bioterrorism



Challenges in Vaccine Production



Manufacturing ChallengesGeneral

• Paradigm = Product defined by Process (live/bacterial vaccines)

• Complex range of vaccine products & sessional variation

• Very diverse production and purification platforms

• Majority of vaccines still produced “traditionally”

• Higher regulatory demands

• Cost for entry is very high

• Risk management

Practical & Technical • Limited number of vaccine CMOs

• Manufacturing: capacity, speed, economy, response

• Capacities not equally distributed worldwide

• Technology knowledge owned by few major accounts

• Analytical assays with low throughput

• Complex impact of process on product

• Time to clinic/approval: PD and clinical material

Picture courtesy of Greg Knobloch



Trends and Strategies



Trends in the Vaccine Market

• New virus based application: immune stimulation, cancer treatment

• ”Old diseases” in formerly ”safe” regions” : Dengue fever, West-Nile virus, Malaria

• Currently many small/mid-size biotech companies pursuing new vaccines

• Differences in regional vaccine & vaccine quality requirements

• Time to market and process economy becoming vital

• New vaccine platform technology: VLP, plants, DNA



Generations of Vaccines

•Killed/ Inactivated

• Subunit

• Toxoid

• Conjugate

• Live-attenuated Viruses

• Adjuvants

• Virus-Like Particles

• Recombinant Antigens

• DNA

• Recombinant vectors

• Novel routes of administration

ImprovedState-of-the-Art Novel



The vaccine landscape is changinge.g. influenza vaccine

System

Lead time

Maturity

Vaccine

Eggs Mammaliancells

Insectcells

No cells

6-9 months ~6 months ~3 months Days ?

On marketNew on market

In 2-4 yearstime

?

All – whole virus, split etc

Molecularapproaches

?

Cell freeEgg based Cell culture based

All – whole virus, split etc



Novel approaches

Pseudomonas fluorescens is a nonpathogenic (BSL-1), Gram-negative, obligately aerobic, bacterium

VaxInnate's fusion vaccine can be efficiently and economically manufactured in E.coli

Tobacco plant-based Proficia™ technologyand its Virus-Like Particles (VLPs).

Virus-like particles expressed in Baculovirus expression system



Efficient Process Development



FDA’s - Guidance for Industry

Process Design:

…Design of Experiment (DoE) studies can help develop process knowledge by revealing relationships, including multifactorial Interactions, between

the variable inputs (e.g., component characteristics or processing parameters) and the resulting outputs (e.g., in-process material, intermediates, or the final product). …

Source: www.fda.gov



Optimize

Plan Predict

PerformOptimize

Evaluate

DoE & HTPD

X2

X1X3-1

1

-1

1-11

Analyze



Screening for hcDNA removalPreDictor™ plates /Capto™ ViralQ

A/Wisconsin/67/2005 (H3N2)A/Solomon Islands/3/2006 (H1N1)



Capto™ ViralQDNA scavenging

0

1000

2000

3000

4000

mAU

0

50

100

150

200

250

mS/cm

0.0 5.0 10.0 15.0 20.0 25.0 30.0 ml

Column: Capto™ ViralQ (CV=1ml)Sample: Influenza A/PR/8/34 (H1N1), produced in MDCK cells

UV_280nm

Cond

DNA level (PicoGreen)

UV_260nm – UV_280nm

Flowthrough=Virus fraction

"Eluate" CIPMDCK DNA levels per dose (45µg HA)

1

10

100

1000

10000

100000

Clarified Harvest UF/DF retentate Capto Q virusfraction

Sucrose grad centr

Purification intermediate

DN

A le

vel (

ng/d

ose)

A/PR/8/34 (inact)A/PR/8/34 A/SI/3/06A/W/67/05



Analytics



Analytical tools - The bottleneck

Typical analytical challenges during viral vector development and production

• 100s of tests per run in both development and production

• Few minutes to hours/days to complete test, particularly lengthy for in-vivo testing

• Varying uncertainty in test accuracy



Analytical methodse.g. in influenza vaccine production

Haemagglutinin (HA) SRID,Dot Blot, HA ELISA (H1N1), HPLC, Western Blot, Hemagglutination assay,

genomic dsDNA qPCR, Molecular Probes PicoGreen™

host cell proteins (HCP) BCA & Bradford protein assay, anti-HCP serum for western blotting, HCP ELISA, 1D-DIGE

virus infectivity TCID50, Fluorescent Focus Assay

quantification



BIACORE assay setup & principle

[Virus standard]( µg/ml)

0

400

800

1200

0 200 400 600

0

0.5

1.0

2.0

4.08.0

x

x

x

xx

x

[ ]( µ

0

400

800

1200

0 200 400 600

0

0.5

1.0

2.0

4.08.0

x

x

x

xx

x

[ ]( µ

Seru

m r

espo

nse

(RU

)

Time (s)

0

400

800

1200

0 200 400 600

0

0.5

1.0

2.0

4.08.0

x

xx

x

xx

x

Sensorgram



HA-assayBIACORE vs. SRID

BIACORE™ SRIDdynamic range

(µg HA/mL) 0.5-10 8-30

sensitivityLOD (µg HA/mL) 0.3 6

LOQ (µg HA/mL) 0.8 13

precision#samples CV < 5%

(%) 97 18



Increasing Productivity



Pandemic PlanningCapacity Time

0

500

1000

1500

2000

2500

mill

ion

dose

s

2006 PandemicPlanning

WHOgoal

• There is a huge gap between the expected demand and the current capacity

• Most countries are looking for in-country domestic solutions

• WHO is driving local capacity build up

20102004

• Need of faster supply – less than 3 months production time

• Currently the manufacturing time is ~6-9 months

• Difficult and time consuming to scale up with old technology



Disposables in vaccine productionProduce faster, safer, and cheaper

Higher project throughput• Shorter setup times• Shorter process times• Less validation efforts/time

Safer operations• Reduced cross-contamination• No re-use, closed system

Reduced costs• Lower capex• Lower start up costs• No cleaning procedures



Vaccine manufacturing solutionsGeneric

TechnologySpecific

SolutionsManufacturing

PlatformsCompleteSolution

Cell harvestingHollow fiber ultrafiltrationNMWC 750kDa, 1 mm lumen

Fermentation

Alkaline lysis

ClarificationNormal flow depth filtration20, 5 and 0.5 µm

ConcentrationHollow fiber ultrafiltrationNMWC 100 or 300kDa

RNA removalGroup separationSepharose 6 Fast Flow

Supercoiled plasmid DNA captureThiophilicaromatic chromatographyPlasmidSelect Xtra

Plasmid DNA polishingAnion exchange chromatographySOURCE 30Q

Concentration/formulationHollow fiber ultra/diafiltrationNMWC 100 or 300kDaSterilizationNormal flow filtration0.2µm

Fill and finish


Fermentation

Alkaline lysis







Fill and finish


Fermentation

Alkaline lysis







Fill and finish

Aseptic Processing

Virus Affinity Media

Chromatography

Filtration

Cell Culture, Bacteria, Plants

Flexible

Conventional

Developing WorldPandemic Solutions



Summary

• DoE or HTPD support PD and optimization (up/downstream) even in a more and more diverse vaccine world

• Technologies like RTP and single-use products can minimize time to market

Variety of existing and future vaccine and virus based applications will put an enormous pressure on vaccine produces, suppliers and academia to work closer together



Thank you

GE Healthcare Bio-Sciences AB, a General Electric company.

GE Healthcare Bio-Sciences ABBjörkgatan 30751 84 UppsalaSweden

ÄKTA, ÄKTAready, Unicorn, Cytodex, WAVE Bioreactor, WAVEPOD, Cellbag, M*Bag, WAVE Mixer, Hot Lips TubeSealer, and LabCrew, Capto, Sephadex, Sepharose, Sephacryl, MAbSelect, FPLC, Source, ReadyToProcess, ReadyMate, ReadyCircuit (pending), ÄKTAcrossflow, Kvick Start, Kvick Lab Packet and BIACORE are trademarks of GE Healthcare companies.

GE, imagination at work and GE monogram are trademarks of General Electric Company.

All third party trademarks are the property of their respective owners.

All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. General Electric Company reserves the right, subject to any regulatory and contractual approval, if required, to make changes in specifications and features shown herein, or discontinue the product described at any time without notice or obligation.

robert morenweiser - dechema · the world vaccine market 93% 7% pharma vaccines 53% 47% 2006 data...

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