Genomic data for alternate production strategies. I. Identification of major contaminating species for Cobalt+2 immobilized metal affinity chromatography

Recent advances in technology have allowed for the identification of complex protein mixtures in a rapid fashion. This report highlights the use of 2D gel electrophoresis, mass spectrometry, and database analysis to determine contaminating species of the Escherichia coli genome that are present duri...

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Bibliographic Details
Published in:Biotechnology and bioengineering 2004-10, Vol.88 (1), p.77-83
Main Authors: Cai, Ying, Moore, Misty, Goforth, Robyn, Henry, Ralph, Beitle, Robert
Format: Article
Language:English
Subjects:
Bacteria
Biological and medical sciences
Biotechnology
Chromatography
Escherichia coli
Fundamental and applied biological sciences. Psychology
Genomics
immobilized metal affinity chromatography
Methods. Procedures. Technologies
Others
proteomics
Various methods and equipments
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Summary:Recent advances in technology have allowed for the identification of complex protein mixtures in a rapid fashion. This report highlights the use of 2D gel electrophoresis, mass spectrometry, and database analysis to determine contaminating species of the Escherichia coli genome that are present during immobilized metal affinity chromatography (IMAC), highlighting Co2+ as the affinity ligand. Four proteins (triosephosphate isomerase, alpha galactosidase, Hsp90, and glucosamine 6‐phosphate synthase) constitute the majority of E. coli proteins that bind and potentially may coelute during chromatography. Results are discussed within the context of changes that when implemented could lead to an increase in IMAC efficiency, not by altering column conditions, but rather by changing the nature of the nuisance proteins that principally reduce column capacity and extend processing times. Such a study illustrates the use of proteome data to aid in bioprocess design. © 2004 Wiley Periodicals, Inc.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.20212