Hong Jin Sung, Azer Nicole, Agarabi Cyrus, Fratz-Berilla Erica J
Center for Drug Evaluation and Research, Office of Product Quality, Office of Biotechnology Products, Division of Biotechnology Review and Research II, U.S. Food and Drug Administration.
Center for Drug Evaluation and Research, Office of Product Quality, Office of Biotechnology Products, Division of Biotechnology Review and Research II, U.S. Food and Drug Administration;
J Vis Exp. 2020 May 14(159). doi: 10.3791/61161.
Primary clarification is an essential step in a biomanufacturing process for the initial removal of cells from therapeutic products within the harvested cell culture fluid. While traditional methods like centrifugation or filtration are widely implemented for cell removal, the equipment for these processes have large footprints and operation can involve contamination risks and filter fouling. Additionally, traditional methods may not be ideal for continuous bioprocessing schemes for primary clarification. Thus, an alternate application using acoustic (sound) waves was investigated to continuously separate cells from the cell culture fluid. Presented in this study is a detailed protocol for using a bench-scale acoustic wave separator (AWS) for the primary separation of culture fluid containing a monoclonal IgG1 antibody from a CHO cell bioreactor harvest. Representative data are presented from the AWS and demonstrate how to achieve effective cell clarification and product recovery. Finally, potential applications for AWS in continuous bioprocessing are discussed. Overall, this study provides a practical and general protocol for the implementation of AWS in primary clarification for CHO cell cultures and further describes its application potential in continuous bioprocessing.
初次澄清是生物制造过程中的一个关键步骤，用于从收获的细胞培养液中初步去除治疗产品中的细胞。虽然像离心或过滤这样的传统方法被广泛用于细胞去除，但这些过程的设备占地面积大，操作可能涉及污染风险和过滤器堵塞。此外，传统方法对于初次澄清的连续生物加工方案可能并不理想。因此，研究了一种使用声波的替代应用，以连续地从细胞培养液中分离细胞。本研究介绍了一种详细的方案，用于使用台式声波分离器（AWS）从CHO细胞生物反应器收获物中对含有单克隆IgG1抗体的培养液进行初次分离。展示了来自AWS的代表性数据，并演示了如何实现有效的细胞澄清和产品回收。最后，讨论了AWS在连续生物加工中的潜在应用。总体而言，本研究为在CHO细胞培养的初次澄清中实施AWS提供了一个实用且通用的方案，并进一步描述了其在连续生物加工中的应用潜力。
