Abstract
An automated two-dimensional chromatographic method has been developed for the isolation and concentration of recombinant fusion proteine with β-galactosidase. The sustem consists of an immunoaffinity column with anti-β-galactosidase antibodies as ligand, followed by an anion-exchange column. It was used for the purification and concentration of recombinant fusion proteins from Mycobbacterium tuberculosis and M. leprae. Small amounts of crude lysates of Escherichia coli were loaded stepwise onto the immunoaffinity column with intermittent washing, elution and re-equilibration. After several cycles the eluate was passed through the anion-exchanger. Using an immunoaffinity gel of 5-ml volume and the anion-exchanger Mono Q HE 5 5 , from 10 ml of crude E. coli lysate (containing up to 50 mg of protein) up to 100 μg of recombinant protein in a 2-ml volume could be isolated overnight.
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