This application note describes a complete process-development workflow, from PreDictor™ plate screening to a 400 mL pilot-scale purification for capturing recombinant pro-insulin expressed in E. coli.
PreDictor 96-well prefilled filter plates, Assist software and Tricorn™ 5/50 columns packed with 1 mL media (resins) were used to develop optimized conditions for the capture step. High-throughput screening quickly narrowed down the number of candidate chromatography media and suggested binding and elution conditions for further investigation. When capture step conditions were further optimized and verified, the purification was scaled up 400-fold.
Scale-up was smooth and predictable, reaching a final-scale yield of 96% and a target molecule purity of 82%. Screening in PreDictor plates also saved considerable time and sample compared with column experiments. A second study describes a similar workflow for the intermediate purification of the insulin obtained following the enzymatic cleavage of pro-insulin.