基于高通量筛选的双特异性抗体纯化方法开发  

Developing a High-Throughput Screening-Based Purification Method for Bispecific Antibodies

作　　者：甘远春 梁忠爽 丁澦[1] 张为杰 GAN Yuanchun;LIANG Zhongshuang;DING Yu;ZHANG Weijie(School of Life Sciences,Fudan University,Shanghai 200433,China;WuXi Biologics(Shanghai FX)Co.,Ltd.,Shanghai 200131,China)

机构地区：[1]复旦大学生命科学学院,上海200433 [2]上海药明生物医药有限公司,上海200131

出　　处：《生物化工》2023年第1期23-29,42,共8页Biological Chemical Engineering

摘　　要：双特异性抗体(BsAbs),简称双抗,是一种能同时结合两个不同靶点或表位的抗体,BsAbs在表达过程中通常伴随产生多种副产物,在纯化工艺中很难被去除。本研究运用高通量筛选技术,通过对SP Sepharose Fast Flow填料的纯化条件进行探索,建立基于Sepharose Fast Flow填料的双特异性抗体纯化方法开发的通用流程,每个筛选条件只需要0.4 mg双抗样品,可同时筛选多达32个条件,筛选过程总耗时2 h,而传统柱层析需耗时64 h。通过高通量迅速筛选方法可以有效去除双抗分子副产物,为解决纯化工艺难题提供了一种新的工艺路线。Bispecific antibodies(BsAbs)are antibodies that can simultaneously bind to two different targets or epitopes.BsAbs are usually accompanied by various by-products,such as homodimers,half antibodies,light chainrelated by-products,and aggregates during the expression process.These by-products show high similarity to the target bispecific antibody and are difficult to remove in the purification process.In this study,using high-throughput screening technology,by exploring the purification conditions of SP Sepharose Fast Flow resin,a general process for developing a bispecific antibody purification method based on Sepharose Fast Flow resin is established.Each screening condition only required 0.4 mg of BsAbs samples can be screened for up to 32 conditions at the same time,and the total screening experiment takes 2 hours,while traditional column chromatography takes 64 hours.Through high-throughput rapid screening method,the molecular by-products of double antibody can be effectively removed,providing a new process route to solve the purification process problems.

关 键 词：双特异性抗体 高通量筛选技术 纯化 SP Sepharose Fast Flow 

分 类 号：Q819[生物学—生物工程]