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首页> 外文期刊>Journal of Biotechnology >Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography
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Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography

机译：使用蛋白A捕获连续逆流切向色谱法从澄清的细胞培养液中纯化单克隆抗体

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摘要

Recent studies using simple model systems have demonstrated that continuous countercurrent tangential chromatography (CCTC) has the potential to overcome many of the limitations of conventional Protein A chromatography using packed columns. The objective of this work was to optimize and implement a CCTC system for monoclonal antibody purification from clarified Chinese Hamster Ovary (CHO) cell culture fluid using a commercial Protein A resin. Several improvements were introduced to the previous CCTC system including the use of retentate pumps to maintain stable resin concentrations in the flowing slurry, the elimination of a slurry holding tank to improve productivity, and the introduction of an "after binder" to the binding step to increase antibody recovery. A kinetic binding model was developed to estimate the required residence times in the multi-stage binding step to optimize yield and productivity. Data were obtained by purifying two commercial antibodies from two different manufactures, one with low titer (similar to 0.67 g/L) and one with high titer (similar to 6.9 g/L), demonstrating the versatility of the CCTC system. Host cell protein removal, antibody yields and purities were similar to those obtained with conventional column chromatography; however, the CCTC system showed much higher productivity. These results clearly demonstrate the capabilities of continuous countercurrent tangential chromatography for the commercial purification of monoclonal antibody products. (C) 2015 Elsevier B.V. All rights reserved.
机译：使用简单模型系统的最新研究表明，连续逆流切向色谱法（CCTC）有可能克服使用填充柱的常规Protein A色谱法的许多局限性。这项工作的目的是优化和实施CCTC系统，以使用市售的Protein A树脂从澄清的中国仓鼠卵巢（CHO）细胞培养液中纯化单克隆抗体。对以前的CCTC系统进行了几项改进，包括使用渗余物泵以保持流动的浆料中稳定的树脂浓度，取消了浆料保持槽以提高生产率，以及将“后粘合剂”引入到粘合步骤中。提高抗体回收率。建立了动力学结合模型以估计在多阶段结合步骤中所需的停留时间，以优化产率和生产率。通过纯化来自两个不同制造商的两种市售抗体获得数据，其中一种具有低效价（约0.67 g / L）和一种具有高效价（约6.9 g / L），证明了CCTC系统的多功能性。宿主细胞的蛋白质去除，抗体产量和纯度与常规柱色谱法相似。但是，CCTC系统显示出更高的生产率。这些结果清楚地证明了连续逆流切向色谱法用于单克隆抗体产品的商业纯化的能力。 （C）2015 Elsevier B.V.保留所有权利。

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