FRACTIONATION OF MURINE H-2 ANTIGENS WITH USE OF DETERGENTS

Three methods were tested for fractionating H-2 antigens from a participate fraction of Sarcoma I using the 51Cr cytotoxic assay to measure antigenic activities. A principal difference among the fractionation procedures was the type of detergent employed to solubilize the antigens. A procedure using Triton X 100 was the least satisfactory. Those using Triton X 114 or cholate were equally effective, but the procedure involving the use of cholate was more convenient. Fractionation of antigens in detergent solutions with (NH 4)2SO 4 was not an efficient procedure. Digestion with trypsin further increased the specific activity of fractions obtained by treatment with detergents. The highest purification attained was a 10-fold increase in activity over that of the starting material. The yield of activity was usually about 60%. No evidence for separation of H-2 antigenic specificities was obtained. Detergent-solubilized preparations were retained wholly or in part on Sephadex and Agarose columns, but these procedures did not result in increased purity. © 1969 by the Williams and Wilkins Co.