Continuous precipitation‐filtration process for initial capture of a monoclonal antibody product using a four‐stage countercurrent hollow fiber membrane washing step

逆流交换 色谱法 降水 错流过滤 化学 过滤(数学) 微滤 脱水 单克隆抗体 材料科学 抗体 生物化学 物理 统计 数学 岩土工程 生物 气象学 工程类 免疫学 热力学
作者
Mirko Minervini,Matthew Mergy,Yuncan Zhu,M.A. Díaz,Craig Pointer,Oleg Shinkazh,Sheldon F. Oppenheim,Steven M. Cramer,Todd M. Przybycien,Andrew L. Zydney
出处
期刊:Biotechnology and Bioengineering [Wiley]
卷期号:121 (8): 2258-2268 被引量:9
标识
DOI:10.1002/bit.28525
摘要

Abstract The significant increase in product titers, coupled with the growing focus on continuous bioprocessing, has renewed interest in using precipitation as a low‐cost alternative to Protein A chromatography for the primary capture of monoclonal antibody (mAb) products. In this work, a commercially relevant mAb was purified from clarified cell culture fluid using a tubular flow precipitation reactor with dewatering and washing provided by tangential flow microfiltration. The particle morphology was evaluated using an inline high‐resolution optical probe, providing quantitative data on the particle size distribution throughout the precipitation process. Data were obtained in both a lab‐built 2‐stage countercurrent washing system and a commercial countercurrent contacting skid that provided 4 stages of continuous washing. The processes were operated continuously for 2 h with overall mAb yield of 92 ± 3% and DNA removal of nearly 3 logs in the 4‐stage system. The high DNA clearance was achieved by selective redissolution of the mAb using a low pH acetate buffer. Host cell protein clearance was 0.59 ± 0.08 logs, comparable to that based on model predictions. The process mass intensity was slightly better than typical Protein A processes and could be significantly improved by preconcentration of the antibody feed material.
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