This webcast features: Sharon Bola, Global Product Manager, Process Chromatography, Bio-Rad Laboratories.
With the increasing global demand for insulin product, there is a need to develop a more efficient and economical process for the production of recombinant insulin therapeutics.
In this webcast, we will present an efficient downstream processing workflow for recombinant human insulin production using Escherichia Coli as the host system. Data will be presented for sample preparation and purification results 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, was used in the capture step of proinsulin and the 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, was found to be highly effective in removing a significant amount of higher molecular weight cleavage product impurities compared to known reverse-phase chromatography steps.
This new purification workflow provides an excellent chromatographic tool for the purification of recombinant human insulin produced from Escherichia Coli inclusion bodies.
- Insulin and proinsulin purification workflow.
- Effects of pH and conductivity.
- Resin characteristics and optimization.
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