ANTIBODY PRODUCTION IN MILK SERUM AFTER VIRUS INSTILLATION OF GOAT MAMMARY GLAND: II. BIOCHEMICAL ISOLATION AND PURIFICATION OF ANTIBODY TO INFLUENZA VIRUS

Abstract

Goat milk was obtained from the right lactiferous sinus following instillation of PR8 influenza type A virus. The proteins precipitated from goat milk serum by (NH4)2SO4 were found to contain antibody activity and further purification was undertaken. In the development of the method the protein solutions were subjected to paper chromatography and paper and polyacetate high voltage electrophoresis, both before and after a number of isolation and purification steps. In addition, preparations from each step were run on the Technicon amino acid analyzer for the presence of amino acids and (or) peptides.As a working procedure, a preparation obtained after cold acetone precipitation was subjected to rivanol treatment, and after electrophoresis a single protein band containing the antibody was obtained. This material was further purified by gel filtration using a Bio-Rad P150 column. The antibody containing protein was finally isolated in the eluate by a fraction collector using phosphate buffer as the eluting fluid. There was excellent recovery of the antibody activity in each preparation.