Downstream Process Development for the Purification of Biosimilar Insulin from Escherichia coli
With the increasing global demand for insulin products, there is a need to develop a more efficient and economical process for the production of recombinant insulin therapeutics. In this webcast, we present an efficient downstream processing workflow for recombinant human insulin production using E. coli as the host system. Data are presented for sample preparation and purification of proinsulin and insulin. Size exclusion was identified as a very efficient method for proinsulin desalting prior to purification. A strong cation exchange resin, Nuvia S Resin, was used in the capture step of proinsulin. Elution in a sodium citrate buffer achieved both good recovery and purity. For the insulin purification step, a high-resolution strong cation resin, Nuvia HR-S Resin, was found to be highly effective in removing a significant amount of higher molecular weight cleavage product impurities, as compared to known reverse-phase chromatography steps. This new purification workflow provides an excellent chromatographic tool for the purification of recombinant human insulin from E. coli inclusion bodies.
Overview
Insulin and proinsulin purification workflow
Effects of pH and conductivity
Resin characteristics and optimization
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