| Abstract: | Two methods of obtaining Fab and Fc fragments from mouse immunoglobulin Gl are described. In the first case the papain protein digest is fractionated on a column with DEAE-or DE-32-cellulose, equilibrated with 0.005 M potassium phosphate buffer, pH 8.0. The Fab fragment is eluted from the column in the starting buffer; the fragment is eluted when the ionic strength of the buffer is increased to 0.4 M. In the second case the protein is fractionated on an ion-exchange resin equilibrated with 0.004 M Tris-H3PO4 buffer, pH 8.5. The whole of the protein applied under these circumstances is bound by the column. The Fab fragment is eluted with 0.04 M Tris buffer containing 0.004 M of a mixture of K-phosphate salts at pH 8.5; the Fc fragment is eluted by increasing the ionic strength by means of phosphate to 0.4 M. Since neither method can yield absolutely pure Fab or Fc fragments, in order to obtain monospecific antisera against these fragments it is necessary to cross-exhaust the antisera with appropriate immunosorbents.Laboratory of Antibody Chemistry and Biosynthesis, N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Sciences of the USSR P. A. Vershilov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 7, pp. 116–119, July, 1979. |
