Parallel, high-throughput purification of recombinant antibodies for in vivo cell assays

被引：15

作者：

Bannister, David ^{[1
]}

Wilson, Andrew ^{[1
]}

Prowse, Leah ^{[1
]}

Walsh, Meg ^{[1
]}

Holgate, Rob ^{[1
]}

Jermutus, Lutz ^{[1
]}

Wilkinson, Trevor ^{[1
]}

机构：

[1] Cambridge Antibody Techol, Cambridge CB1 6GH, England

来源：

BIOTECHNOLOGY AND BIOENGINEERING | 2006年 / 94卷 / 05期

关键词：

scFv; purification; IgG; His Tag; phage display; IMAC; cation exchange chromatography;

D O I：

10.1002/bit.20914

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We describe a method for high-through put, parallel purification of secreted proteins to analyse large numbers of protein samples in cell-based assays for the discovery of protein therapeutics. The procedure 'is generic and capable of 96 parallel purifications and compatible, in both yield and purity, with a wide assay range. By optimising expression and purification steps as well as using novel hardware, in particular a chromatography press capable to purify target proteins from viscous media, we exemplify the process for the generation of single-chain Fv antibody fragments (scFv) and the purification of full-length IgG. The described process can operate robustly with a throughput of over 2,000 samples per month. (c) 2006 Wiley Periodicals, Inc.

引用

页码：931 / 937

页数：7

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