Continuous precipitation for monoclonal antibody capture using countercurrent washing by microfiltration
Continuous precipitation for monoclonal antibody capture using countercurrent washing by microfiltration
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DOI：
10.1002/btpr.2886
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发表时间：
2019-08-08
影响因子：
2.9
通讯作者：
Zydney, Andrew L.
中科院分区：
文献类型：
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作者：
Li, Zhao;Gu, Qin;Zydney, Andrew L.
人们对使用沉淀作为集成连续下游过程的一部分用于高价值治疗性蛋白质的初始捕获的可能性重新产生了兴趣。目前在大多数细胞培养工艺中实现的高产物滴度极大地促进了沉淀，这与在高滴度下性能降低的色谱工艺形成鲜明对比。本研究使用可逆交联（氯化锌，ZnCl2）和体积排阻（聚乙二醇）试剂的组合，使用连续管式沉淀反应器直接从收获的细胞培养液中沉淀单克隆抗体产物。然后将沉淀物脱水并使用切向流过滤连续洗涤，其中使用逆流分级配置来减少所需的洗涤缓冲液的量并增加宿主细胞蛋白质的去除。通过在临界滤液通量以下操作膜组件以避免结垢来实现长期操作。实验结果证明了这种完全连续的集成沉淀过程在实验室规模上的可行性，设计计算用于探索影响该系统初始抗体捕获性能的关键因素。
There is renewed interest in the possibility of using precipitation for initial capture of high-value therapeutic proteins as part of an integrated continuous downstream process. Precipitation is greatly facilitated by the high product titers now achieved in most cell culture processes, in sharp contrast to chromatographic processes whose performance is reduced at high titers. The current study used a combination of reversible cross-linking (zinc chloride, ZnCl2) and volume exclusion (polyethylene glycol) agents to precipitate a monoclonal antibody product directly from harvested cell culture fluid using a continuous tubular precipitation reactor. The precipitates were then dewatered and continuously washed using tangential flow filtration, with a countercurrent-staged configuration used to reduce the amount of wash buffer required and increase host cell protein removal. Long-term operation was achieved by operating the membrane modules below the critical filtrate flux to avoid fouling. Experimental results demonstrate the feasibility of this fully continuous integrated precipitation process at bench scale, with design calculations used to explore the key factors affecting the performance of this system for initial antibody capture.