Synthesis of Recombinant Human Insulin

Data of Diabetic Patient

Since Banting & Best invented insulin hormone in 1921, amount of diabetic patients increased worldwide.Their glucose level increased due to disorder of insulin production treatment by insulin injection from animal pancreas

Insulin is produced and secreted by beta cells of Langerhans pancreas, that regulates the use and storage of carbohydrates.

Problem?

Animal insulin swine and cow

Human insulin slight different in composition

As a consequence : human body produces antibodies against the animal insulin inflamationat the area of injection

Long-term complication occurred

Decreasing of animal insulin production

Structure of Insulin

Chemically, insulin is a small and simple protein , consists of 51 amino acids, 30 construct 1 polypeptide chain and 21 amino acids construct the second chains. Both chains are linked by disulfide bond

Synthesis of human insulin using E.coli

Human Insulin is produced by inserting gene of insulin at suitable vector of Escherichia coli(production of Humulin, Eli Lily) by recombinant DNA Technology

Selectively obtaining insulin producing beta epithelioid cells

from a pancreas, a small circular DNA in a bacteria, called plasmid, modified

by inserting human DNA gene for a

precursor proinsulin

Plasmid containing proinsulin gene is then transformed into Eschericia coli

Dispersing cells in a nutrient amino acid-rich medium composition in an open aeration incubation vessel maintained at about 32° - 39°C

Protein formed is partly B-galactosidase, on chain A and chain B of the insulin A and B are extracted from fragments of B-galactosidase and purified

Separating the cells from the media,inactivebacteria by heat sterilization, and extracting genomic material from the cells,

Proinsulin harvested by cutting by

restriction enzyme to produce human

insulin

Further Process is centrifugation and eliminating

unused cells. Purification is carried out by

liquid chromatography

and crystallization