Integrated Clarification and Purification of Monoclonal Antibodies by Membrane Based Separation of Aqueous Two-Phase Systems.

Thomas Kruse,Markus Kampmann,Jochen Strube,Axel Schmidt

doi:10.3390/antib8030040

Thomas Kruse, Markus Kampmann + Show 2 more

Open Access

https://doi.org/10.3390/antib8030040

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Abstract

Therapeutic monoclonal antibodies (mAb) are used for the treatment of numerous serious diseases, which have led to an increasing demand over the last decades. Increased cell density and mAb titer of the cultivation broth lead to great challenges for the subsequent clarification and capture operations in the downstream process. As an alternative approach to the conventional downstream process, a selective mAb extraction via an aqueous two-phase system (ATPS) directly from the cultivation broth of a mAb producing industrial relevant chinese hamster ovary (CHO) cell line was investigated. An efficient purification of the mAb was accomplished by the ATPS composition. The phase separation was realized by a newly developed membrane based phase separator. Moreover, a complete cell removal was integrated into this process by the used membrane. A selectivity between both phases was achieved by membrane modification. Yields up to 93% in the light phase and removal of process related impurities were obtained after aqueous two-phase extraction (ATPE). Phase separation performance as well as contact angles on the membrane were characterized for different ATPS. ATPE directly from the cultivation broth in combination with the new membrane based phase separation led to a mAb yield of 78% with a simultaneous reduction of deoxyribonucleic acid (DNA) and host cell protein (HCP) load.

Highlights

Due to their enormous importance as active pharmaceutical ingredients for the treatment of numerous severe diseases like immunological disorders [1,2], cancer [2], and inflammatory as well as infectious diseases [1,2], the demand for monoclonal antibodies is rising [3]
aqueous two-phase extraction (ATPE) directly from the cultivation broth in combination with the new membrane based phase separation led to a monoclonal antibodies (mAb) yield of 78% with a simultaneous reduction of deoxyribonucleic acid (DNA) and host cell protein (HCP) load
Phase separation was accomplished by the use of a modified hydrophobic membrane with surfactants

Summary

Introduction

Due to their enormous importance as active pharmaceutical ingredients for the treatment of numerous severe diseases like immunological disorders [1,2], cancer [2], and inflammatory as well as infectious diseases [1,2], the demand for monoclonal antibodies (mAb) is rising [3]. Improved media and feeding strategies in the upstream process of mAb producing cell lines, like chinese hamster ovary (CHO) cells, have resulted in significantly increased cell density and titers up to 25 g/L [4,5,6]. These achievements have led to high challenges for the subsequent clarification and capture operations, causing a bottleneck in the downstream process (DSP) [7]. As subsequent mAb capture step protein A affinity chromatography is used in most platform processes, offering the advantages of high yield and purity as well as a volume reduction [8,12]

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Conclusion