Functional polymer affinity matrix for purifying hexahistidine-tagged recombinant protein.
A functional polyacrylic acid (PAA) adsorbent has been prepared for metal chelate affinity chromatography. It has been found to chelate nickel ion Ni2+ strongly, and was evaluated for the ability to bind proteins containing neighbouring histidine residues. The principle of the technique was illustrated with Aeromonas hydrophila outer membrane protein OmpTS. DNA elements coding for adjacent histidines were fused to the Aeromonas hydrophila ompTS gene. Subsequent expression in E. coli resulted in the production of hybrid protein His6-OmpTS that could be purified by Ni2+-PAA affinity chromatography. The remarkable specificity found makes it an attractive addition to the range of adsorbents for metal chelate affinity chromatography.
Zeng
Q
Q
Key Laboratory for Polymeric Composite and Functional Materials of Ministry of Education, Material Science Institute, Zhongshan University, Guangzhou, China.
Xu
J
J
Fu
R
R
Ye
Q
Q
D000327:Adsorption; D000333:Aeromonas / Q000737:chemistry; D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D001483:Base Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D011108:Polymers / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013055:Spectrophotometry, Infrared
D001425:Bacterial Outer Membrane Proteins; D011108:Polymers; D011994:Recombinant Proteins; D004247:DNA