Performance optimization of continuous countercurrent tangential chromatography for antibody capture.
Performance optimization of continuous countercurrent tangential chromatography for antibody capture.
复制标题
用于抗体捕获的连续逆流切向色谱的性能优化。
DOI：
10.1002/btpr.2250
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发表时间：
2016
影响因子：
2.9
通讯作者：
Shinkazh,Oleg
中科院分区：
文献类型：
--
作者：
Dutta,AmitK;Tan,Jasmine;Napadensky,Boris;Zydney,AndrewL;Shinkazh,Oleg
最近的研究表明，连续逆流切向层析（CCT）可以有效地从澄清的细胞培养液中纯化单克隆抗体。CCTC具有克服传统填充床蛋白A色谱的许多局限性的潜力。本文从产品收率、杂质去除、总生产率和缓冲液使用等方面探讨了优化的方法。建模基于实验室规模工艺开发和工业合作伙伴提供的两种含单克隆抗体的澄清中国人卵巢细胞培养原料的蛋白A捕获的CCTC实验数据。评估了树脂结合能力和动力学以及分期策略和缓冲液循环的影响。发现结合步骤中的最佳分级提供了更好的产率，并将整个系统的生产率提高了8- 16%。在洗涤和洗脱步骤中使用更高的阶段数可导致缓冲液使用量显著降低（减少约40%）以及杂质去除量增加（去除量增加约200 log）。通过在洗涤和再生步骤中回收缓冲液，可以进一步减少缓冲液的使用量（约35%）。使用较小粒径树脂的初步结果表明，由于结合步骤中传质限制的减少，CCTC系统的生产率可提高2.5倍，高达190 g mAb/L树脂/hr。这些结果提供了一个坚实的框架，设计和优化的捕获应用程序的CCTC技术。© 2016美国化学工程师学会Biotechnol.程序：32：430-439，2016
Recent studies have demonstrated that continuous countercurrent tangential chromatography (CCTC) can effectively purify monoclonal antibodies from clarified cell culture fluid. CCTC has the potential to overcome many of the limitations of conventional packed bed protein A chromatography. This paper explores the optimization of CCTC in terms of product yield, impurity removal, overall productivity, and buffer usage. Modeling was based on data from bench‐scale process development and CCTC experiments for protein A capture of two clarified Chinese Hamster Ovary cell culture feedstocks containing monoclonal antibodies provided by industrial partners. The impact of resin binding capacity and kinetics, as well as staging strategy and buffer recycling, was assessed. It was found that optimal staging in the binding step provides better yield and increases overall system productivity by 8–16%. Utilization of higher number of stages in the wash and elution steps can lead to significant decreases in buffer usage (∼40% reduction) as well as increased removal of impurities (∼2 log greater removal). Further reductions in buffer usage can be obtained by recycling of buffer in the wash and regeneration steps (∼35%). Preliminary results with smaller particle size resins show that the productivity of the CCTC system can be increased by 2.5‐fold up to 190 g of mAb/L of resin/hr due to the reduction in mass transfer limitations in the binding step. These results provide a solid framework for designing and optimizing CCTC technology for capture applications. © 2016 American Institute of Chemical EngineersBiotechnol. Prog., 32:430–439, 2016