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Parallel, high‐throughput purification of recombinant antibodies for in vivo cell assays
Author(s) -
Bannister David,
Wilson Andrew,
Prowse Leah,
Walsh Meg,
Holgate Rob,
Jermutus Lutz,
Wilkinson Trevor
Publication year - 2006
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20914
Subject(s) - recombinant dna , throughput , high throughput screening , chromatography , yield (engineering) , in vivo , chemistry , antibody , affinity chromatography , cell , target protein , computational biology , biochemistry , biology , computer science , gene , materials science , enzyme , microbiology and biotechnology , immunology , telecommunications , metallurgy , wireless
We describe a method for high‐throughput, 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. © 2006 Wiley Periodicals, Inc.