变速上样在MabSelect填料上分离WLB303

doi:10.13523/j.cb.20160609

中国生物工程杂志

2016, Vol. 36

Issue (6): 65-75 DOI: 10.13523/j.cb.20160609

技术与方法

变速上样在MabSelect填料上分离WLB303

蒙国基, 邓义熹, 李乐, 罗豪晖, 于玉根

深圳万乐药业有限公司深圳 518029

Promotion in ProteinA Chromatography of WLB303 Monoclonal Antibody by Using Dual Flowrate to Load Sample

MENG Guo-ji, DENG Yi-xi, LI Le, LUO Hao-hui, YU Yu-gen

Shenzhen Main Luck Pharmaceuticals Inc., Shenzhen 518029, China

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摘要：

随着抗体表达量的提升和生产规模的扩大,Protein A亲和层析不仅需要高载量填料,也需要提高工艺效率。变速上样的方法可以在满足载量要求的同时大大缩短工艺耗时。通过测定WLB303单克隆抗体在GE MabSelect填料多个保留时间的动态载量,建立一元三次方程拟合载量和保留时间的关系。以该方程计算获得填料在快、中、慢速下不同保留时间的动态载量表,并以此作为变速上样组合的参考依据。在2.7ml层析柱上使用精纯样品测试最快的变速上样组合的可行性;然后用常规纯化工艺在7ml层析柱上使用细胞培养澄清液,对最快的变速上样组合和恒速上样的两种工艺周期进行了比较,证实变速上样的方式能明显提升整体工艺效率。

关键词： 变速上样; 单克隆抗体; 动态载量; ProteinA亲和层析; 工艺效率

Abstract:

With the advances in cell culture expression levels and production capacity in the last two decades,it is crucial to improve the binding capacity of protein A resin and decrease processing time. Then the dual flowrate loading strategy is proposed. A cubic equation of GE MabSelect dynamic binding capacity(DBC) was created by measuring several DBC values of the retention time with WLB303 mAb. A unique DBC table of GE MabSelect was generated subsequently to design dual flowrate loading strategy. The fastest dual flowrate loading mode was firstly confirmed by using purified WLB303 mAb to practice on a 2.7 ml column. And it was demonstrated to show obvious promotion in process efficiency by using clarified harvest to practice on a 7 ml column.

Key words: Dual flowrate Monoclonal antibodies ProteinA chromatography Dynamic binding capacity Process efficiency

收稿日期: 2015-11-19 出版日期: 2016-01-26

ZTFLH:

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通讯作者: 蒙国基 E-mail: mengguoji@wanle.com.cn

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引用本文:

蒙国基, 邓义熹, 李乐, 罗豪晖, 于玉根. 变速上样在MabSelect填料上分离WLB303[J]. 中国生物工程杂志, 2016, 36(6): 65-75.

MENG Guo-ji, DENG Yi-xi, LI Le, LUO Hao-hui, YU Yu-gen. Promotion in ProteinA Chromatography of WLB303 Monoclonal Antibody by Using Dual Flowrate to Load Sample. China Biotechnology, 2016, 36(6): 65-75.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160609 或 https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I6/65

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