Streamlined purification of recombinant Alpha-1 antitrypsin from CHO cell culture using integrated chromatography within a single unit operation

Abstract

Recombinant Alpha-1 Antitrypsin (A1AT) produced from CHO cell culture is a promising alternative to the commercially available drug, which is derived from the limited supply of blood plasma. By developing and optimizing the purification strategy through the combination of appropriate chromatography mechanisms and buffer conditions, we successfully achieved end-to-end purification of A1AT from cell culture supernatant to the final drug substance leveraging integrated chromatography within a single unit operation, with quality comparable or even superior to commercial drugs. With comparable processing time and buffer consumption, the proposed approach—integrating 2 anion exchange chromatography (AEX) and 1 hydrophobic charge induction chromatography (HCIC) —achieved 99.9 % purity and 27 ppm host cell protein (HCP), compared to 96.9 % purity and 2860 ppm HCP using the conventional method with a single AEX column in batch mode. Compared to the conventional approach of using one column per unit operation, this streamlined process can increase the productivity of A1AT by reducing unit operations and eliminating the need for sample holding and conditioning between chromatographic steps, thereby potentially reducing the manufacturing cost of recombinant A1AT produced from CHO cell culture.