A General Method for the Specific Purification of Antiprotein Antibodies

Abstract

A procedure has been developed which is probably of general applicability for the specific purification of antiprotein antibody. The principle involved is the alteration of the specific antigen so that it can be separated from the antibody brought down in the specific precipitate. The alteration was accomplished by coupling the specific antigen with diazotized arsanilic acid. It was shown that such an antigen can be removed from an alkaline solution by a suspension of calcium aluminate while rabbit serum antibody remains in solution under these conditions. The coupled antigen retained the ability to precipitate specifically the antiprotein antibody. The purified antibody was obtained from the specific precipitate with the coupled antigen by dissolving the washed precipitate in calcium hydroxide solution and removing most of the antigen with a suspension of calcium aluminate, leaving purified antibody in solution. By this method, solutions of antibody to bovine serum albumin and to bovine serum globulin have been obtained with essentially all of the protein precipitable as specific antibody. The yields were about 30 per cent.