INSIghT PhARMA REPORTSA Division of Cambridge Healthtech Institute

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therapeutic protein production: a changing landscape examines the implications of and advances in upstream and downstream operations used in the production of therapeutic proteins. Improvements in protein yields per unit volume, the significance of introducing disposables throughout the process and the shift to animal extracts-free culture media are three of the major themes covered by this report. The impact of global competition, lower barriers to entry and the shift in the production bottleneck from upstream to downstream unit operations are presented together with likely shifts impacting the production outlook over the next five years. More than a dozen tables and figures in this report summarize the report’s findings.

Some of the topics highlighted in the report:• Pros and cons of animal cell culture• Development of alternative production cell lines• Improving protein production per volume of culture media• Impact of disposables on cost and availability of proteins• Bottlenecks appear on the downstream side of production• Shift away from capital spending on fixed, reusable infrastructure• Tackling biosimilar regulatory questions• Dimensions of Asian and EU competition

A ChAnging LAndsCApeK. John Morrow, Jr., PhD

July 2010

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O v E R v I E w

Therapeutic Protein Production: A Changing Landscape Author: K. John Morrow, Jr., PhD

Bioprocessing is the branch of biotechnology dealing with the production and purification of biological materials of commercial interest. This report focuses on the wide range of disciplines applied to the rapid and economical production of protein molecules. While most antibody biologics are anti-cancer agents, there are a number of other disease indications, including immune dysfunction, infectious disease and mental illness management, fueling the growth of this therapeutic area.

Trends in the upstream and downstream areas of monoclonal antibody production are encouraging and should help control the manufacturing costs of monoclonal antibodies. As indicated in this report, an increase in the use of disposables in upstream and downstream unit operations is underway. This shift away from fixed, reusable equipment is driving change in the industry, making available to cash-strapped small companies workable product quantities at affordable costs and allowing companies to enter clinical trials with promising but higher-risk, second-tier candidates. The availability of disposables has caused major biotech companies to question their strategy of large capital investments in reusable equipment.

This report identifies the continuing upstream versus downstream gap problem for the industry. Progress toward producing more protein per unit volume of culture medium has moved more rapidly and is placing pressure on the rates at which these proteins can be purified at the downstream end. This overall expansion has motivated regulatory agencies in Europe to redefine intellectual property guidelines and rules governing biosimilars. Legislation governing biosimilars has been passed in the US as part of the recent 2010 health care legislation.

This report also evaluates rising concern over the threat of Asian and European competition to the American biotech industry.

ABOuT ThE AuThOR

K. John Morrow, Jr., PhD, is

a writer and consultant for

the biotechnology industry.

He obtained his PhD in

genetics from the University

of Washington in Seattle,

and completed his training

with post-doctoral studies

in Italy at the Universitá di

Pavia and Philadelphia at the

Fox Chase Cancer Institute.

He has held faculty positions

at the University of Kansas

and at Texas Tech University

Health Sciences Center. His

writings include over 200

peer-reviewed journal papers,

non-peer–reviewed coverage

of the biotechnology industry,

books, and marketing reports.

A number of companies,

including Meridian Bioscience,

Affitech, Henderson-Morley

Biotechnology, Brandwidth

Communications, and Emergent

Technologies have taken

advantage of his consultancy

services, provided through

Newport Biotechnology

Consultants. He resides in

Newport, KY.

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Number (and Percentage Values Siding the Bars) of Recombinant Proteins Approved as Biopharmaceuticals in Different Production Systems

Source: Microb Cell Fact. 2009;8:17.

INSIghT PhARMA REPORTSA division of Cambridge Healthtech Institute

250 First Avenue • Suite 300 • Needham, MA 02494 • 781-972-5444 • InsightPharmaReports.com

Executive Summary

Chapter 1Introduction1.1 Bioprocessing in the 21st Century

A Word of CautionState of the Art

What is Bioprocessing?An Industry Committed to Mammalian Cell Expression Systems (Golden 2009, See Appendix)

A Changing LandscapeThe PipelineAntibodies Rule the RoostCorporate StrategiesPressures and Opportunities

1.2 Bioprocessing Advances Are Incremental in Scope

1.3 Range of the Industry Challenge

Chapter 2Upstream Processing2.1 Mammalian Cell Culture Expression Systems

Historical Background Media Development and the Evolution of Therapeutic AntibodiesCell Line Development: Engineering for Better Performance

Developing Overproducing Cell LinesMedarex Selection Procedure to Avoid RoyaltiesConventional Cell Lines: CHO, NSO, PER.C6, Other TypesChinese Hamster Ovary (CHO) Cells NSO CellsPer.C6 Cells (Golden 2009, See Appendix)Human Cell LinesSelection for Overproducers

2.2 Mammalian Cell Culture Media Introduction to Mammalian Cell Culture Media Strategies Elimination of Serum From Mammalian Cell Culture Media

Protein HydrolysatesOnline Sources of InformationMany Options for Cell Culture Media Optimization

2.3 Post-Translational ModificationsGlycosylationS-GlutathionylationOther Modifications

2.4 Other Production AlternativesBacteriaYeastGreen Leafy Plants

Chlamydomonas reinhardtii BioLexis; DuckweedMicroalgae

Insect CellsChesapeake PERL, Inc.Alternatives to Baculovirus for Insect Cell Protein Synthesis

Protozoan Expression Hosts

2.5 Bioreactors: Design and FunctionComputer Modeling of Bioreactor PerformanceDisposable Bioreactor ComponentsDisposables and Rapid Response

Chapter 3Downstream Processing3.1 Disposables, Reusables

Disposable Technology Comes of AgeDisposable Storage OptionsThe Disposable ConundrumThe Future of Disposables

3.2 Purification StrategiesFine-Tuning Contaminant Removal Through Filtration Technology

Current Polishing Strategies: The ToolboxSingle-Pass Tangential Flow Filtration (TFF)Recombinant Polyclonal Purification ChallengesFilter Reconfiguration for Better Antibody Purification

Affinity Tag Purification (Culp 2009 and Tomaselli 2009, See Appendix)

Self-Cleaving Affinity Tag Purification Protein AOther Protein A AlternativesTrends in Bioprocessing from the European Union

3.3 Monitoring Tools: HPLC-MS

Chapter 4Optimizing Bioprocess Development4.1 Time is Money

Avoiding Revenue Loss The Next Challenge The Example of Insulin

Develop Processes That Will Survive Upscaling Economic Savings From DisposablesFlexibility Generates Cost Savings

Lean, Not Necessarily Mean

Chapter 5Current Regulatory Status of Biologics5.1 The Biologics Problem

European Guidelines on BiologicsCanadian Regulatory Policy for BiologicsStatus of US Biosimilar Regulation

Chapter 6Conclusions Drawn While on the Road Ahead;

Thoughts on the Future of Bioprocessing6.1 Predicting the Future

Small, Incremental Steps Will Guide Progress in Bioprocessing

6.2 The Rise of Foreign Competition China’s Advance

Drawbacks to China’s AdvanceChinese Regulatory PolicyUS and China: Price CompetitionWill the US Throw Away Its Advantages in Bioprocessing?

IndiaThe European ThreatCMOs: Upsides and Downsides

6.3 Marketing Issues; CostsAre Revolutionary Developments Possible in Bioprocessing?Disposables in the Ascendant

6.4 Options and CaveatsGood News and Challenges AheadShort-Term Trends in BioprocessingR&D is a Major Cost Driver

F I g u R E S & TA B L E S

TABLESTable 1.1 A Wide Range of Species Under Study for

Protein ProductionTable 2.1 Selected List of Approved Recombinant

Therapeutic Proteins Produced in Mammalian Cell Lines

Table 2.2 Mammalian Cell Culture Products and Media

Table 2.3 Recommendations by Regulatory Agencies and Physicians‘ Organizations for the Avoidance of Human or Animal Proteins in Drug Manufacturing

Table 2.4 Approved Therapeutic Protein-Based Products from E. coli

Table 2.5 Comparison of Plants Built to Accommodate Reusable Versus Disposable Upstream Bioprocessing Facilities

Table 3.1 Various Polishing OptionsTable 3.2 Typical Platform for MAb PurificationTable 3.3 Companies Dealing with Liquid

Chromatography Systems and ProductsTable 4.1 FDA Biologics Approvals, 2009Table 6.1 Comparative Salaries (in US $$) for High-

Tech Occupations in Various Countries, 2008

FIguRESFigure 2.1 Number (and Percentage Values Siding the

Bars) of Recombinant Proteins Approved as Biopharmaceuticals in Different Production Systems

Figure 2.2 General Scheme for Commercial Development and Production of a Plasma-Free Recombinant Therapeutic Protein. In Every Step of the Manufacturing Process, Animal- or Human-Derived Additives are Replaced by Plant-Based or Synthetic Molecules.

Figure 6.1 Cell-Free Synthesis Production Example

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R E L AT E D R E P O R TMonoclonal Antibodies:Pipeline Analysis and Competitive Assessment

In the nearly 35 years since the first process for creating mAbs was introduced, they have remained a centerpiece of the growing biotechnology industry. Thirty therapeutic mAbs have been approved around the world, including 23 in the United States. A number of these drugs have attained blockbuster status, with sales reaching the coveted billion-dollar mark and well beyond. Rituxan, Remicade, Avastin, Herceptin, and Humira alone generated sales of over $4 billion each in 2008, and global sales for this entire sector surpassed $30 billion last year.

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R E L AT E D C O N F E R E N C ECambridge Healthtech Institute’s Tenth Annual

January 10-14, 2011Hotel Del Coronado | San Diego, CALeading the way from Discovery through Development.

Celebrate CHI’s 10th Annual PepTalk event! This week long event features strong scientific programming comprising fourteen confer-ences within topic focused pipelines. The four distinct pipelines range from applying protein discovery research, to developing downstream protein expression, characterization, formulation, and production that ultimately lead to clinical applications.

• Pipeline 1: Formulating Biologics

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• Pipeline 4: Highter-Throughput Protein Expression

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