Plant Design

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Plant Design
insulin hexamer
plant
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Insulin

• insulin is hormone which regulates glucose level
  in blood
• produced by islets of Langerhans in pancreas
• deficiency results in diabetes

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Discovery

• Frederick Banting and Charles Best (1922)
• Eli Lilly, Indiana first commercial production
  (monopoly)
• Nordisk Insulin Lab (1923) joined with Novo
• Denmark became largest insulin producer outside
  USA

Banting/Best lab at UofT
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Traditional production

• pancreas from pigs or cows
• crystallized from pancreatic tissue extracts with
  addition of zinc
• gel filtration to separate proteolytic enzymes,
  insulin dimers, proinsulin
• ion exchange to further remove contamiinants

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Human versus Porcine

• human insulin differs from porcine insulin by 1
  amino acid (threonine at position 30 the
  carboxyl-terminus) in human B-chain replaced by
  alanine in pig insulin

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Porcine to human

• porcine insulin converted to human insulin by
  trypsin digestion
• cuts B-chain between 22 and 23 and between 29 and
  30
• synthetic octapeptide is inserted (amino acids 23
  to 30)

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Recombinant human insulin

• FDA approved in 1982 as first approved
  recombinant biopharmaceutical
• E. coli (intracellular) or Saccharomyces
  cerevisiae (extracellular)
• GRAS (generally regarded as safe)
• Nova Nordisk, Eli Lilly, Hoechst (now Aventis)
• cells produce insulin precursor

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Recombinant insulin
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Our process
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Downstream steps

• vertical tank and centrifuge, remove cells
• precursor purified on cation exchanger (washed
  and eluted with acid)
• diafilter concentrates precursor
• converted to insulin ester with transpeptidase
  (residual denatured precursor)
• Reverse Phase-HPLC isolates insulin ester

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purification steps

• second diafilter
• further purified on anion exchanger
• ester hydrolyzed with acid in WM reactor to
  produce human insulin
• second RP-HPLC, eluted with acetonitrile
• concentrated with ultrafilter
• insulin crystallized and separated from
  acetronitrile with centrifuge
• insulin crystal, amorphous insulin freeze-dried

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Feeds and Products (kg)

• Feed ammonia (320) glucose (3800) air
  (28,000) water (183,000) purified water for
  downstream (211,000)
• Output biomass (1400) CO2 (2200) ammonia (80)
  water (2300)
• Product stream insulin crystal (190) insulin
  (1.4) water (0.6)

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Your job should you choose to accept it

• undertake design of assigned unit
• consideration of process alternatives
• complete specifications
• supplier(s), cost, sizing
• outline theory and principle of operation
• engineering drawings of process configuration