High Throughput Development of Non-Protein A Monoclonal Antibody Purification Process using Mini-Columns and Bio-Layer Interferometry

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Pall-Downstream-2014High throughput (HT) sorbent screening is widely used for developing purification process saving time and sample volume. This study describes the development of a non-protein A purification process for a monoclonal antibody (MAb) expressed in a CHO cell supernatant. For the capture step, 54 process conditions were screened in three days using 200 µL mini-columns packed with five different sorbents. Capacity and yield were evaluated using Bio-Layer Interferometry (BLI) with protein A biosensors and MAb purity, using SEC-HPLC. A 340-fold scale-up of the best capture conditions was conducted using a lab-scale column packed with MEP HyperCelTM sorbent. The eluate of the first step was used for screening the intermediate purification step on mini-columns in a bind/elute mode. A total of 94 process conditions were tested on five sorbents in seven days using a design of experiment (DoE) approach, leading to the selection of CM Ceramic HyperD® sorbent. After the two chromatography steps, the MAb’s purity was 97% with a yield of 90%. A F/T polishing step will  remove remaining impurities.

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