The Isolation and Characterization of an Anti-Sheep Red Blood Cell Antibody Having Limited Heterogeneity

A rabbit (RB1) was induced to produce anti-sheep red blood cell antibodies of limited heterogeneity in abnormally high concentrations. These antibodies were isolated by a new procedure which involves the cross-linking of stroma with glutaraldehyde to produce a reusable immunosorbent. This immunosorbent permitted the isolation of a pure antibody preparation, primarily of the IgG class. The small amount of IgM antibodies present was removed by Sephadex chromatography to yield a pure IgG product (99.7%) which gave a discrete dense band by cellulose acetate electrophoresis and three or four bands by isoelectric focusing. Also, by isoelectric focusing, one major and one minor light chain band and two major heavy chain bands were observed. Moreover, all of the antibody molecules were of the same allotype and had a characteristic idiotypic specificity.