Purification of hemagglutinin from Haemophilus paragallinarum using monoclonal antibody.

The purification of hemagglutinin from Haemophilus paragallinarum was attempted using affinity chromatography with monoclonal antibody. The antigen eluted from the affinity column using potassium thiocyanate buffer agglutinated chicken erythrocytes. In immunoblotting of the eluted antigen, a single band with monoclonal antibody was found as well as the crude antigen. When the chickens were immunized with the eluted antigen, they produced the hemagglutination inhibition (HI) antibody, and they showed protection against challenged exposure with H. paragallinarum strain 221. These results indicated that the HA antigen of H. paragallinarum was a protective antigen.