Identification and characterization of native proteins of Escherichia coli BL-21 that display affinity towards Immobilized Metal Affinity Chromatography and Hydrophobic Interaction Chromatography Matrices.
The purpose of this study was to identify and characterize Escherichia coli proteins which display affinity towards both Immobilized Metal Affinity Chromatography (IMAC) and Hydrophobic Interaction Chromatography (HIC). Co(II) IMAC was chosen as the primary capture step, followed by HIC employing different concentrations of salt to promote adsorption. Results provided insight on this rather small pool of E. coli proteins. Nine out of the ten have isoelectric values less than six, and half are considered nonessential. These data indicate that the combination of IMAC and HIC could be developed as a potent method for the purification of recombinant proteins by judicious choice of the salt concentration used to promote HIC, the development of E. coli strain(s) deficient in certain genomic proteins, and the design of an IMAC-HIC affinity tail for recombinant protein isolation based on the very proteins deleted from the genome.
Tiwari
Neha
N
Ralph E. Martin Department of Chemical Engineering, 3202 Bell Engineering, University of Arkansas, Fayetteville, AR 72701, USA.
Woods
Lauren
L
Haley
Ryan
R
Kight
Alicia
A
Goforth
Robyn
R
Clark
Kevin
K
Ataai
Mohammad
M
Henry
Ralph
R
Beitle
Robert
R
D002845:Chromatography / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000302:isolation & purification; D057927:Hydrophobic and Hydrophilic Interactions; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*
D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins