The capture of recombinant antibodies from cell culture broth is the first critical step of downstream processing. We were able to develop a precipitation‐based method for the capture and purification of monoclonal antibodies based on divalent cations, namely ZnCl2. Traditional precipitation processes have to deal with high dilution factors especially for resolubilization and higher viscosity due to the use of PEG as precipitation or co‐precipitation agent. By the use of the crosslinking nature of divalent cations without the use of PEG, we kept viscosity from the supernatant and resolubilization dilution factors very low. This is especially beneficial for the solid–liquid separation for the harvest and wash of the precipitate in continuous mode. For this harvest and wash, we used tangential flow filtration that benefits a lot from low viscosity solutions, which minimizes the membrane fouling. With this precipitation based on ZnCl2, we were able to implement a very lean and efficient process. We demonstrated precipitation studies with three different antibodies, Adalimumab, Trastuzumab, and Denosumab, and a continuous capture case study using tangential flow filtration for precipitate recovery. In this study, we achieved yields of 70%.

中文翻译：

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无需聚乙二醇，通过 ZnCl 2 沉淀连续捕获重组抗体

从细胞培养液中捕获重组抗体是下游处理的第一个关键步骤。我们能够开发出一种基于沉淀的方法，用于捕获和纯化基于二价阳离子（即 ZnCl2）的单克隆抗体。传统的沉淀过程必须处理高稀释因子，特别是由于使用 PEG 作为沉淀剂或共沉淀剂而导致再溶解和更高的粘度。通过利用二价阳离子的交联性质而不使用 PEG，我们将上清液的粘度和再溶解稀释因子保持在非常低的水平。这对于以连续模式收集和洗涤沉淀物的固液分离特别有利。对于这种收获和洗涤，我们使用了切向流过滤，该过滤从低粘度溶液中受益匪浅，从而最大限度地减少了膜污染。通过这种基于 ZnCl2 的沉淀，我们能够实施非常精简且高效的工艺。我们演示了使用三种不同抗体（阿达木单抗、曲妥珠单抗和地诺单抗）的沉淀研究，以及使用切向流过滤进行沉淀回收的连续捕获案例研究。在这项研究中，我们实现了 70% 的产率。