Purificaiton by Surcrose Density Gradient Zonal Centrifugatin and Affinity Coloum Chromatogrphy of Antigenic Ssubstancaes of Antigenic Susbstances from the Livers and Mice Infected with Tyzzer's Diseae

Antigenic substances from livers of mice infected with Tyzzer's disease were purified by means of sucrose density gradient zonal centrifugation and affinity column chromatography using antiserum and checking antigenicity with the complement fixation test. Fractions obtained from zonal centrifugation fell into three main groups with different molecular weights, two of which (Fr. I and Fr. 11) positively reacted with antiserum in the complement fixation test. Both fractions were further purified by affinity column chromatography. The molecular weights of the main antigenic substances derived from Fr. I and Fr. I1 were determined to be about 52000 and 66000, respectively, by means of SDS-PAGE.