"An overview of technology

spects of biotechnology industry"

Venkata Ramana

January 19, 2007

1
Contents

1. Introduction

2. Recombinant DNA technology

3. Product development

4. Life cycle of biotechnology product

5. Summary

2
Contents

1. Introduction

2. Recombinant DNA technology

3. Product development

4. Life cycle of biotechnology product

5. Summary

3
Birth of biotech industry
Stanley Cohen Herbert Boyer Robert Swanson

Genentech was founded in 1976 by Boyer and Swanson

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 Biopharmaceuticals Market Size

US$ bn

140 125
120 2,862
2,503 3,565
100
4,150 8,426
80 50
60
40 25
15 4,400
20 1,703
5,731
0
1999 2001 2005 2010
Blood Factors Erythropoetins Hormones Interferons
Insulin MAbs Growth Factors Others
 Present biopharma penetration in global
pharma market (US$392 billion) is 8%.
 It is projected to reach about 17% of the Biopharmaceutical market by Segments (2002)
projected global pharma market of Total Market Size - US$ 33 billion
US$750 billion by 2010.
Source: Reuters Business Insight
(All figures in US$ billions)

Global biopharmaceuticals market has grown at a CAGR of

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Biotechnology products

•Natural or modified proteins produced using rDNA technology

•Antibodies against receptors

•Subunit vaccines

•Fusion proteins

•Gene therapy products

•Chimeric proteins

•si RNA products

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Recombinant proteins
 Microbial recombinant proteins
 Interferon alpha
 Growth hormone

 Mammalian recombinant proteins

 Interferon beta
 Erythropoietin

 Monoclonal antibodies
 Anti EGFR2 (Herceptin)
 Anti CD20 (Rituxan)

Recombinant proteins are fastest growing health care

products 7
Contents

1. Introduction

2. Recombinant DNA technology

3. Product development

4. Life cycle of biotechnology product

5. Summary

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Macromolecules present in a living organism

 DNA/RNA (information flow)

 Proteins (Building blocks)

 Lipids (Part of cell membrane and involved in various

functions)

 Carbohydrates (Primary source of energy and storage)

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 Central dogma of life
Transcription Translation
Duplication DNA RNA PROTEIN

Central dogma is still valid even after 50 years

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Restriction enzymes

 First restriction enzyme was isolated from Haemophilus

influenzae (1970)
- Bacterial enzymes
- Hydrolyze DNA
 Names are derived from the bacterial strain
 Basic tools of gene cloning

Restriction enzymes and indispensable tools

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Restriction enzymes
Werner Arber Daniel Nathans Hamilton Smith

Nobel prize for the discovery of restriction enzymes and

their application to problems of molecular genetics 12
 Standard molecular biology techniques

•Gene isolation

•Polymerase chain reaction (PCR)

•Restriction and modification enzymes

•Transformation

•Screening

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Standard molecular biology techniques

Gene isolation

•Genomic DNA

•mRNA

•Synthetic gene

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 Standard molecular biology techniques

•Polymerase chain reaction (PCR) Denaturation

•DNA template
Annealing
•Primers

•DNA polymerase
Extension

•Thermal cycler

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Making recombinant DNA

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DNA digestion and ligation

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Transformation

Blue white
selection

Medium containing antibiotic,

IPTG and X-gal

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Recombinant DNA technology

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Hybridoma (MAb) technology

White: Mouse Black: Human

Chimeric Antibody Humanized antibody

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Expression in various hosts

•Bacteria (E.coli)
•Yeast (Saccharomyces, Pichia)
•Chinese hamster ovary cells
•Insect cells
•Mouse cells
•Plants
•Transgenic animals

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Contents

1. Introduction

2. Recombinant DNA technology

3. Product development

4. Life cycle of biotechnology product

5. Summary

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Bioprocess flow sheet
Cultivation medium Frozen working seed
preparation & (recombinant E. coli)
Sterilization
Inoculum (pre culture) preparation

Production (Fermentation)

Cell separation
Biomass
(Centrifugation)

Cell homogenisation Inclusion body (IB) purification IB solubilization

Refolding

Purification
(Chromatography)

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Recombinant protein purification

1 IB 3 AEX

Protein Protein
5 GFC

2 HIC 4 CONC Protein

Protein
Protein

More than 98% pure protein was obtained after purification 24

Purity analysis of protein

Protein

SDS-PAGE gel – Silver stained

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MALDI-TOF of tryptic digest of a protein

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Mass of intact protein by mass spectrometry

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 Biopharmaceutical facility

Special facilities are needed for the production of

biopharmaceuticals 28
Contents

1. Introduction

2. Recombinant DNA technology

3. Product development

 Life cycle of biotechnology product

 Summary

29
Biopharmaceuticals: Discovery to the market

GLP

GMP

Research
Discovery
(Research) Process Development
Preclinical testing & IND
(Lab and animal testing - Toxicity) Manufacturing
Phase I Clinical trial
(20-30 healthy volunteers – Safety & Dosage)
Phase II Clinical trial
(100-300 patient volunteers – Efficacy & Side effect)
Phase III Clinical trial
(1000-5000 patient volunteers – long term effect)

BLA, FDA review and approval

Postmarketing testing

0 2 4 6 8 10 12 14 16
Year

Patent life of a biotech product is typically 20 years from

the date of filing 30
Source of products for a company

•Licensing

•Acquisition of a company owning the IP

•In house discovery

Licensing is a major route for companies to start the

product development 31
Life cycle of a biotech product
 Development of new variants
Long and slow acting insulins

 Competition from small molecules

Gleevec replaced interferon alpha therapy for CML

 Better serum half lives

Pegylation of interferons and GCSF

 Lower toxicity
Replacement of interleukins with monoclonal antibodies

New products are being developed using existing backbone

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Life cycle of a biotech product –
New technologies
 Monoclonal antibodies

 si RNA

 Gene therapy

 Novel delivery of proteins

 Fusion proteins

 Chimeric proteins

New technologies are making a mark on the life cycle of a

product 33
Life cycle of a biotech product-
New antibodies

 Mouse monoclonal antibodies

 Chimeric antibodies

 Humanized antibodies

 Human antibodies

More than 70% of new products under development are

antibodies 34
Contents

1. Introduction

2. Recombinant DNA technology

3. Product development

 Life cycle of biotechnology product

 Summary

35
Summary

 Effective health care products are being developed using

recombinant DNA technology
 New technologies are helping in developing better drugs
replacing existing products
 Regulation on biosimilars will lead to affordable biotech
products

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Thank You