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

Autor: Bannister D; Cambridge Antibody Technology, Granta Park, Milstein Building, Cambridge CB1 6GH, UK. david.bannister@cambridgeantibody.com, Wilson A, Prowse L, Walsh M, Holgate R, Jermutus L, Wilkinson T
Jazyk: angličtina
Zdroj: Biotechnology and bioengineering [Biotechnol Bioeng] 2006 Aug 05; Vol. 94 (5), pp. 931-7.
DOI: 10.1002/bit.20914
Abstrakt: 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.
((c) 2006 Wiley Periodicals, Inc.)
Databáze: MEDLINE
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