Application of an aqueous two‐phase systems high‐throughput screening method to evaluate mAb HCP separation

• DOI: 10.1002/bit.22900
• Publication Date: 2010-08-17T12:06:31Z
• Authors: Stefan A. Oelmeier; Florian Dismer; Jürgen Hubbuch
• Keywords: Biopharmaceutical; Aqueous two-phase system

Aqueous two-phase systems (ATPSs) as separation technique have regained substantial interest from the biotech industry. Biopharmaceutical companies faced with increasing product titers and stiffening economic competition reconsider ATPS an alternative to chromatography. As implementation of is material, time, labor intensive, a miniaturized automated screening process would be beneficial. In this article such method, its statistical evaluation, application biopharmaceutical task are shown. To speed up early stage profiling cloud-point method for binodal determination was developed. PEG4000-PO(4) binodals were measured automatically manually found identical within experimental error. The procedure applied model system industrial task. at protein concentration 0.75 mg/mL used. influence pH, NaCl addition, tie line length investigated. Lysozyme protein, two monoclonal antibodies, host cell pool yield partition coefficients determined values lysozyme. antibodies shifted bottom into upper phase by addition NaCl. This shift occurred lower when pH closer pI distributed protein. Addition NaCl, increase in PEG4000 led significant loss mAb due precipitation. Capacity limitations these thus demonstrated. chosen allowed reduction 50% HCP recovery greater than 95% target proteins. might not industrially relevant compared current chromatographic procedures, developed allows fast evaluation more suitable optimized given