The Purification of Antibody to PR8 Influenza A Virus

Abstract

The physico-chemical procedures a t present available for the separation of proteins appear to possess inadequate resolving power for the isolation of specific antibodies from mixtures such as the y-globulin fraction of human plasma [23, 3, 141. The failure t o achieve significant purification by these means is due presumably to the similarity of the various antibodies in solubiIity, size, shape, net charge and other physical chemical properties. A different approach takes advantage of the natural interaction of an antibody with its homologous antigen t o separate the specific antibody from the mixture [8, 9, 261. The complex is then isolated and dissociated. The recovery of antibody is facilitated by linking the antigen to a solid phase [2, 131. Cellulose or ion exchange resins, which offer several means of forming insoluble complexes witd the antigen, have proved particularly useful. The difficulty remains, however, of achieving satisfactory dissociation of antibodies from protein antigens. In such cases good yields of antibody can be obtained only b y use of extremely acid or alkaline reactions.The present study has been directed primarily a t overcoming this difficulty. Working on the hypothesis that the formation of the antigen-antibody complex results in part from electrostatic interaction between the molecules, the dissociating action of a synthetic polyelectrolyte has been studied. Polymethacrylic acid for instance has been shown t o interact with proteins depending on the relative

Intravenous Administration of Human γ-Globulin

“…(3) y-globulin, modified either by enzymatic or acid treatment, has a similar antibody spectrum as standard y-globulin and causes no reaction in contrast to the nonmodified standard preparation [10].…”