Continuous precipitation-filtration process for initial capture of a monoclonal antibody product using a four-stage countercurrent hollow fiber membrane washing step.
Continuous precipitation-filtration process for initial capture of a monoclonal antibody product using a four-stage countercurrent hollow fiber membrane washing step.
复制标题
使用四阶段逆流中空纤维膜洗涤步骤初始捕获单克隆抗体产品的连续沉淀过滤过程。
DOI：
10.1002/bit.28525
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发表时间：
2023
影响因子：
3.8
通讯作者：
Zydney,AndrewL
中科院分区：
文献类型：
--
作者：
Minervini,Mirko;Mergy,Matthew;Zhu,Yuncan;GutierrezDiaz,MarioA;Pointer,Craig;Shinkazh,Oleg;Oppenheim,SheldonF;Cramer,StevenM;Przybycien,ToddM;Zydney,AndrewL
产品滴度的显著增加，加上对连续生物加工的日益关注，重新引起了人们对使用沉淀作为蛋白A色谱法的低成本替代品用于单克隆抗体（mAb）产品的初级捕获的兴趣。在这项工作中，使用管式流沉淀反应器从澄清的细胞培养液中纯化商业相关的mAb，通过切向流微滤提供脱水和洗涤。使用在线高分辨率光学探针评价颗粒形态，提供整个沉淀过程中粒度分布的定量数据。在实验室构建的2级逆流洗涤系统和提供4级连续洗涤的商业逆流接触滑动装置中获得数据。该过程连续运行2 h，mAb总产率为92 ± 3%，4阶段系统中的DNA去除率接近3 log。通过使用低pH乙酸盐缓冲液选择性再溶解mAb实现高DNA清除。宿主细胞蛋白清除率为0.59 ± 0.08 log，与基于模型预测的结果相当。工艺质量强度略优于典型的蛋白A工艺，并且可以通过抗体进料材料的预浓缩来显著改善。
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.