The use of separated subunits of yeast phosphofructokinase for the isolation of subunit-specific antibodies from polyclonal antisera.

The subunits alpha and beta of yeast phosphofructokinase have been separated under denaturing conditions by ion exchange chromatography and then separately immobilized on Sepharose 6MB. The resulting affinity gels were used to fractionate polyclonal phosphofructokinase antisera from rabbit into subunit-specific antibodies. In addition to subunit-specific antibodies also antibody fractions which recognize both types of subunits of phosphofructokinase could be desorbed from the affinity gels. Antibodies directed either to the alpha- or to the beta-subunits were found to inhibit the enzyme activity. The results are discussed in terms of a partial identity of the alpha- and beta-chain of phosphofructokinase.