synthesis of recombinant human insulin - download
TRANSCRIPT
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
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,