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 during immobilized metal affinity chromatography (IMAC), highlighting Co(2+) 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.
Cai
Ying
Y
Department of Chemical Engineering, University of Arkansas, 3202 Bell Engineering Center, Fayetteville, Arkansas 72701, USA.
Moore
Misty
M
Goforth
Robyn
R
Henry
Ralph
R
Beitle
Robert
R
D000595:Amino Acid Sequence; D002614:Chelating Agents; D002846:Chromatography, Affinity / Q000379:methods*; D003035:Cobalt; D030541:Databases, Genetic; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000235:genetics*; D013058:Mass Spectrometry; D008670:Metals; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011506:Proteins / Q000235:genetics; D040901:Proteomics / Q000379:methods*; D013045:Species Specificity
D002614:Chelating Agents; D008670:Metals; D011506:Proteins; D003035:Cobalt