A high-throughput purification of monoclonal antibodies from glycoengineered Pichia pastoris

被引：14

作者：

Jiang, Youwei ^{[1
]}

Li, Fang ^{[1
]}

Button, Michelle ^{[1
]}

Cukan, Michael ^{[1
]}

Moore, Renee ^{[1
]}

Sharkey, Nathan ^{[1
]}

Li, Huijuan ^{[1
]}

机构：

[1] GlycoFi Inc, Lebanon, NH 03766 USA

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2010年 / 74卷 / 01期

关键词：

Pichia pastoris; Monoclonal antibody; High-throughput; Purification; Fermentation; Glycosylation; GLYCOSYLATION; EXPRESSION; PROTEINS; YEAST;

D O I：

10.1016/j.pep.2010.04.016

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Glycoengineered Pichia pastoris provides a unique platform for screening monoclonal antibody (mAb) leads and high expressing strains. A simple, economic, and high-throughput purification for mAb from P. pastoris fermentation has been developed that can be easily operated in various commercially available liquid handlers. The method includes the use of STREAMLINE rProtein A in a 96-well platform and demonstrates good linear alignment and reproducibility in a wide concentration range. The antibody titers measured by the method have less than 15% variation in comparison to spiking titers. The mAb titer and quality obtained from this method are comparable to that from conventional column chromatography. The method can process hundreds of expression screening samples in a day, not only to accurately determine titers, but also to generate milligram quantities of mAb for quality assessment, including purity, folding, glycosylation, and antigen binding affinity. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：9 / 15

页数：7

共 20 条

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