Studies on human antibodies. IV. Purification and properties of anti-A and anti-B obtained by absorption and elution from insoluble blood group substances.
Insoluble blood group substances prepared by copolymerization of soluble blood group substances with N-carboxy-L-leucine anhydride were used to absorb blood group antibodies from two human, high-titered anti-A sera. After the absorbants were washed free of nonspecific serum proteins, blood group antibodies were eluted either with pH 3.62 acetate buffer, or at neutral pH with monosaccharide haptens of the A or B antigenic determinants (N-acetyl-D-galactosamine or D-galactose respectively). The purified anti-A antibodies were characterized, immunoelectrophoretically, as gammaM-, gammaA-, and gammaG-immunoglobulins. These were further separated into gammaM- and gammaG-fractions by gel filtration or density gradient centrifugation. Both gammaM- and one of the two gammaG-antibody fractions contained K and L light chain determinants; the remaining gammaG-fraction was comprised, almost totally, of type K molecules. Precipitability of the purified anti-A immunoglobulins by blood group A substance varied from 43 to 89%. The agglutinating activity per unit N of the isolated gammaG-anti-A was found to equal, in one case, and to exceed, in the second, that of the gammaM-antibodies from the same individuals. The marked differences between gammaM- and gammaG-antibody fractions in quantitative hapten inhibition studies were interpreted to mean that the antibody-combining site of the isolated eluted gammaG-anti-A was significantly larger than that of the eluted gammaM-anti-A. Whether these data connote differences in combining site size between entire immunoglobulin classes in an individual serum or simply reflect the properties of highly selected antibody populations cannot be stated at present.
D000818:Animals; D000906:Antibodies; D000937:Antigen-Antibody Reactions; D001789:Blood Group Antigens; D005260:Female; D005752:Gastric Mucins; D006801:Humans; D007122:Immunoelectrophoresis; D066298:In Vitro Techniques; D010051:Ovarian Neoplasms; D011233:Precipitin Tests; D013552:Swine; D005719:gamma-Globulins
D000906:Antibodies; D001789:Blood Group Antigens; D005752:Gastric Mucins; D005719:gamma-Globulins