抄録
By means of affinity chromatography anti-catalase antibody was isolated and purified from sera of rabbits immunized with purified rat liver catalase. The antigen was conjugated with CNBr-activated Sepharose 4B. IgG fraction of the immune sera was subjected to chromatography on a column of this immunoadsorbent. The antibody was released from the antigen at pH 2.8-3.0.
Antibody thus purified, was estimated to have a purity over than 90%. It showed an identical behavior on electrophoresis to the rest of IgG of the immune serum, and its amino acid composition was extremely similar to that of the latter. One μg of the purified antibody was found to be equivalent to 0.4μg of the antigen, suggesting that no inactivation occurred during the course of purification.
Diffusion of rat liver cell fractions in agar against the purified anti-catalase antibody gave a second weaker precipitin line in addition to the predominant one, which was common to purified catalase.
Iodination of the purified antibody yielded a preparation of I125-antibody having a specific radioactivity of 400, 000 cpm per mg, with a slight loss in immunological activity.
