Recombinant protein production and Continuous Process Verification (CPV)

Abstract

Biotherapeutic Protein production has largely been driven by bacterial systems, which remains most attractive due to low cost, high productivity, and its rapid use. Inexpensive and ease of cultivation makes the Escherichia coli one of the most broadly used bacterial hosts for the production of intracellular recombinant human biotherapeutic protein. The genetics of E.coli are far better characterized than those of any other microorganisms. The influence of different carbon sources on growth of recombinant Escherichia coli and recombinant human biotherapeutic protein production in cytoplasmic space was studied in fermentor. This project involves the use of E.coli to produce the recombinant protein. The aim of this project was to increase the yield and purity of desired protein. This was done by optimization of pH, temperature and agitation. It was initially carried out at shake flask level. The scale up process was carried out considering one factor at a time approach (OFAT), by either maintaining pH at X± 0.5 or temperature at Y± 0.50 C at a time. In this batch fermentation process, the induction was done using IPTG (Isopropyl β-D-1-thiogalactopyranoside) and the parameters were maintained till the end of the fermentation process. The batch was continued till 13.5 hrs of fermentation and then centrifuged to separate the biomass pellet from broth and lysis was done to obtain IB (Inclusion bodies).