Growth of Inductrial Fermentation Field

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Growth of Inductrial Fermentation Field

• Phase I Biblical ? 1900
• Beer, wine, pickles, cheese, alcohol
• Phase II 1900 ? 1973
• Solvents, organic acids, antibiotics, amino
  acids, vitamins, enzymes, insecticides,
  nucleotides, coccidiostats, gums, alkaloids,
  plant growth factors, animal growth promotants,
  SCP, steroids
• Mutation, screening, biochemical engineering
• Phase III 1973 ? ?
• Genetic engineering

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If I were you, Id buy Cetus stock right now
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• Taq polymerase is the most popular of all
  reagents requested on NIH grants
• (Dalton, 1999)

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BIOPHARMACEUTICALS

• 2002 market 31 billion 8 of 390 billion
  drug market
• 2003 market 40 billion
• 2004 market 55 billion 10 of 550 billion
  drug market
• 197 products on market
• Clinical entries
• 1995 91 1998 114 2004 800
• 1996 100 1999 132
• 1997 119 2000 137
• Fastest growing Monoclonal Antibodies
• Mainly for cancer
• Market 3- 4 billion
• 13 products
• 160 in clinic
• 2002 FDA approved 20 new products and 15 new
  indications
• (Robinson, 2002 Mullin, 2003,2004 Sandman,
  2005 Maggon, 2005)

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BIOTECH MARKET
(Burrill, 1999 Cunningham, 2002 Gupta Roy,
2002 Mullin, 2004 Schmidt, 2004 Maggon, 2005
Sundman, 2005)
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RECOMBINANT HUMAN INSULIN

• 1994 Market 600 million
• 1995 Market 700 million
• 1997 Market 1 billion
• 2001 Market 2.9 billion

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EPO MARKETS

• 1990 200 million
• 1992 410 million
• 1993 600 million
• 1994 1 billion
• 1995 2 billion
• 1999 3 billion
• 2000 3.5 billion
• 2001 5.7 billion
• 2002 8 billion
• 2004 11.8 billion

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• EPO was fortunate choice for AMGENs first
  product.
• 2004 market 11.8 billion (all companies)
• Market evaluation in 2002 of Amgen 60 billion
• Larger than Pharmacia-Upjohn (50 billion)
• Larger than Schering-Plough (50 billion).
• Approaching Eli-Lilly, Abbott and Wyeth (80-90
  billion each).
• (Bowden and Crow, 2002 Thayer,
  2002 Maggon, 2005)

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RECOMBINANT G-CSF MARKET

• 1991 230 million
• 1993 719 million
• 1995 936 million
• 1999 1.3 billion
• 2001 1.5 billion
• 2004 3.0 billion

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• Human Growth Hormone (HGH)
• Used to be made from pituitary glands of
  cadavers
• Expensive
• Often contaminated with prions, transmitting
  Creutzfeld-Jakob disease
• Recombinant HGH from E. coli does not cause
  prion disease
• Market in 2004 1.8 billion
• (BioPharm Guide 2000 Maggon, 2005)

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Human Growth Hormone

• Expected Sales10 million/year
• 1989 Market 120 million
• 1990 Market 200 million
• 1991 Market 300 million
• 1993 Market 700 million
• 1998 Market 1.1 billion
• Japan 417 million
• USA 335 million
• Europe 332 million
• 1999 Market 1.4 billion
• 2002 Market 1.7 billion
• 2004 Market 1.8 billion

(Pranik, 1999 DePalma, 2003 Maggon, 2005)
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Factor VIII

• Blood clotting protein, deficient in
  hemopheliacs.
• Conventional treatment administration of blood
  coagulant products from human plasma.
• 60 of hemopheliacs have been infected with HIV,
  hepatitis or other diseases.
• Approved in 1992.

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Interferons

• Alpha-
• 70-90 remission in hairy cell leukemia
• 60-70 remission in chronic myeloid leukemia
• 20-40 remission in Kaposis Sarcoma
• Also used for Hepatitis B and C and genital warts
• 1989 world market 125 million
• 1992 world market 800 million
• 1996 world market 1 billion
• 2002 world market 1.4 billion
• Gamma-
• Approved for chronic granulomatous disease in
  1990
• Beta-
• Approved for multiple sclerosis in 1993.
• 1995 world market 225 million
• 2002 world market 1 billion

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Monoclonal Antibodies

• In 2000, the largest group of biopharmaceuticals
  in industrial development 750 by 250
  organizations 35 in phase III clinical trials.
• 3 4 billion market in 2003
• Marketed products
• Cancer
• Heart disease
• Transplant rejection
• Early mouse products unsuitable
• Antigenic response
• Had to be humanized by removal of certain
  peptide fragments
• Made in transgenic mice
• Made in human immune cells

(Sedlak, 2000 Baez et al., 2000 Matthews, 2003)
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MONOCLONAL ANTIBODIES (mAbs)

• Previously, polyclonal antibodies (pAbs) were
  used
• Produced by entire immune system of an animal
• Contained various specificities, various
  affinities, very variable.
• mAbs developed by Georges Kolter and Cesar
  Milstein in UK in 1975 by fusing normal
  antibody-producing spleen cells from mice with
  tumor cells to yield long-lived cell lines
  (hybridomas).
• Awarded Nobel Prize in 1984 along with Niels
  Jerne for his theoretical contribution.
• Produced by single cells of immune system
• Mice treated with a single antigen
• Allowed to show immune response
• Spleens removed
• Cells are fused with cells of mouse lymphoma cell
  line
• Hybridoma cells cloned and screened
• Isolation of cells excreting specific antibody
• Cells humanized by genetic engineering
• By 2002 20 approved (market - 4 billion), 100in
  clinical trials
• Bring a drug or radioisotope to a target protein
  and bind it.
• (C. Coty, 2002 A. Loffet, 2002 Pizzi, 2003)

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BIOCONVERSIONS

• Becoming essential to the fine chemical industry
  due to the demand by pharmaceutical and
  agrochemical companies for single-isomer
  intermediates.
• (Rogers, 1999)

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NEW ENZYME ACTIVITIES BY DIRECTED EVOLUTION

• ß-glucosidase ? ß-galactosidase
• ß-glucuronidase ? ß-galactosidase
• Lipase ? phospholipase
• Indole-3-glycerolphosphate synthetase ?
  phosphoribosylanthranilate isomerase
• (H. Zhao et al., 2002)

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WHOLE GENOME SHUFFLING

• Increased acid-tolerance of commercial lactic
  acid producing Lactobacillus
• Tylosin
• Conventional Eli Lilly strain improvement
  examining 1 million survivors gave 6-fold
  increase over 20 years

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WHOLE GENOME SHUFFLING (cont.)

• WGS in 96 well plates
• mutation
• Wild-type 11 strains
  (1.2-1.3x wt.)
  22,000 survivors protoplast
  fusion
  1000
  fusants protoplast fusion
  7 strains (4-9x
  wt.) 7 strains (2-3x wt.)
• 1000 fusants
  

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WHOLE GENOME SHUFFLING (cont.)

• Top strain provided 25 more than best
  conventional strain.
• Time 1 year
• Total strains 24,000
• (R. Paitnak et al., 2002 Y-X. Zhang et al., 2002)

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