Downstream Processing

Reduced Cost: Fewer steps in downstream processing reduce the overall cost of the manufacturing process. Increased Yield: Fewer steps prevent product loss during processing, which would improve overall product yield. Time Efficiency: Less steps in processing save time, as it means fewer procedures and lower processing time.

Fermentation Broth: E. coli} containing the recombinant plasmid (carrying the gene for human insulin, “Humulin") is cultured in a fermenter. Cell Harvest and Lysis: After the fermentation is complete, the bacterial cells are harvested and lysed to release all the intracellular contents including the Humulin protein. Cell debris removal and protein solubilization: Cell debris is removed by methods like centrifugation and filtration. The Humulin protein needs to be solubilised by using buffers, before it can be separated from other proteins. Chromatography: The solubilized protein mix is then passed through chromatographic columns to purify the protein using different methods based on size, charge or affinity. Refolding and purification: The protein can be purified and refolded to reach its active form. Final Purification: The resulting product may undergo additional steps of purification to meet the pharmaceutical standards.

The purification step which involves ‘Solvent Extraction/Chromatography’ is highly metabolite-specific because it uses chemical properties of a compound to purify it. Both solvent extraction and chromatography are methods for separating or purifying certain specific chemical compounds from complex mixtures.