Abstract

A functional polyacrylic acid (PAA) adsorbent has been prepared for metal chelate affinity chromatography. It has been found to chelate nickel ion Ni 2+ 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 His 6-OmpTS that could be purified by Ni 2+–PAA affinity chromatography. The remarkable specificity found makes it an attractive addition to the range of adsorbents for metal chelate affinity chromatography.

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