PURIFICATION OF RECOMBINANT ALPHA-AMYLASE BY IMMUNOAFFINITY CHROMATOGRAPHY WITH ANTIPEPTIDE ANTIBODY

Adsorption characteristics of an anti-peptide antibody, obtained by im munization of eight amino acids in the C-terminal region of chimeric a lpha-amylase of rice alpha-amylase isozymes, were studied by use of th e chimeric enzyme and the peptide used for immunization. This anti-pep tide antibody adsorbed the enzyme, as well as the peptide antigen, wit h sufficient affinity for immunoaffinity purification and was used for purification of the enzyme secreted from yeast cells. Chimeric a-amyl ase was purified by immunoaffinity chromatography to high purity in on e step from the fermentation broth. One-third of the secreted enzyme w as not adsorbed by the column of anti-peptide antibody because of proc essing in the C-terminal region.