Automated high throughput microscale antibody purification workflows for accelerating antibody discovery

被引:18
作者
Luan, Peng [1 ]
Lee, Sophia [1 ]
Arena, Tia A. [2 ]
Paluch, Maciej [3 ]
Kansopon, Joe [1 ]
Viajar, Sharon [1 ]
Begum, Zahira [4 ]
Chiang, Nancy [1 ]
Nakamura, Gerald [1 ]
Hass, Philip E. [3 ]
Wong, Athena W. [2 ]
Lazar, Greg A. [1 ]
Gill, Avinash [1 ]
机构
[1] Genentech Inc, Dept Antibody Engn, San Francisco, CA 94080 USA
[2] Genentech Inc, Dept Early Stage Cell Culture, San Francisco, CA 94080 USA
[3] Genentech Inc, Dept Prot Chem, San Francisco, CA 94080 USA
[4] Genentech Inc, Purificat Dev, San Francisco, CA 94080 USA
关键词
antibody; automation; high throughput; hybridoma; liquid handling system; purification; rat IgG2a; small scale; tip column; transient expression; PHARMACEUTICAL-INDUSTRY; CHROMATOGRAPHY;
D O I
10.1080/19420862.2018.1445450
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
To rapidly find best-in-class antibody therapeutics, it has become essential to develop high throughput (HTP) processes that allow rapid assessment of antibodies for functional and molecular properties. Consequently, it is critical to have access to sufficient amounts of high quality antibody, to carry out accurate and quantitative characterization. We have developed automated workflows using liquid handling systems to conduct affinity-based purification either in batch or tip column mode. Here, we demonstrate the capability to purify >2000 antibodies per day from microscale (1 mL) cultures. Our optimized, automated process for human IgG1 purification using MabSelect SuRe resin achieves similar to 70% recovery over a wide range of antibody loads, up to 500 mu g. This HTP process works well for hybridoma-derived antibodies that can be purified by MabSelect SuRe resin. For rat IgG2a, which is often encountered in hybridoma cultures and is challenging to purify via an HTP process, we established automated purification with GammaBind Plus resin. Using these HTP purification processes, we can efficiently recover sufficient amounts of antibodies from mammalian transient or hybridoma cultures with quality comparable to conventional column purification.
引用
收藏
页码:624 / 635
页数:12
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