Functional polymer affinity matrix for purifying hexahistidine-tagged recombinant protein

A functional polyacrylic acid (PAA) adsorbent has been prepared for metal c helate affinity chromatography. It has been found to chelate nickel ion Ni2 + strongly, and was evaluated for the ability to bind proteins containing n eighbouring histidine residues. The principle of the technique was illustra ted with Aeromonas hydrophila outer membrane protein OmpTS. DNA elements co ding for adjacent histidines were fused to the Aeromonas hydrophila ompTS g ene. Subsequent expression in E. coli resulted in the production of hybrid protein His(6)-OmpTS that could be purified by Ni2+-PAA affinity chromatogr aphy. The remarkable specificity found makes it an attractive addition to t he range of adsorbents for metal chelate affinity chromatography. (C) 2001 Elsevier Science B.V. All rights reserved.