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US 7,608,583 B2Granted
Process for the extraction and isolation of insulin from recombinant sources
AssigneeWockhardt Limited
InventorsMaharaj K Sahib, Edupuganti B Raju, Umesh S Shaligram
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Abstract
The present invention relates to the extraction and isolation of insulins from recombinant sources, particularly those expressed in and secreted by yeasts. Organic solvents have been used to extract host surface bound forms of insulin polypeptide. In addition, procedures for combining the steps medium clarification, solvent extraction and chromatography, in order to effect the simultaneous isolation and purification of soluble and membrane bound forms of insulin, is disclosed.
Claims8
A process for enhanced recovery of recombinant insulin comprising:
- (a) expressing the insulin in yeast, wherein the expressed insulin is secreted into the culture medium;
- (b) adding a water miscible organic solvent to the fermentation culture broth, wherein the fermentation culture broth contains the secreted insulin and the yeast, wherein the water miscible organic solvent is selected from the group consisting of ethanol, isopropanol, t-butanol, acetic acid, dimethylformamide, dimethylsulfoxide, acetonitrile, dioxan, ethylene glycol, and propylene glycol, and wherein the water miscible organic solvent optionally contains additives;
- (c) clarifying the fermentation culture broth containing the added water miscible organic solvent by centrifugation and decanting the supernatant from the pellet;
- (d) isolating the insulin from the supernatant.
1 more element
Dependencies are inferred from the claim text.