Separation of Cells According to Surface Antigens by the Use of Antibody-Coated Columns. Fractionation of Cells Carrying Immunoglobulins and Blood Group Antigen

It has been found possible to separate cells according to their surface anti gens by the use of antibody-coated columns. High efficiency columns were made by double-layer principles, first coating beads with antigen followed by antibody in excess. Such columns could be shown to contain a high amount of free antigen-binding sites for the relevant antigens. Lymphoid cells were thus fractionated according to their surface concentration of immunoglobu lin and a highly selective retention of mouse B lymphocytes was observed when filtering spleen cells through an anti-immunoglobulin column prepared according to the above procedure. No evidence of retention of mouse T lymphoid cells was observed in the same system. By the use of anti-gamma-2a immunoglobulin columns, it was found possible to deplete a population from memory cells potentially capable of synthesizing gamma-2a antibodies. No evidence was found that columns prepared in the described manner would function through combining with receptors on lymphoid cells for antibody-antigen complexes. By using anti-A blood group columns, it was possible to selectively retain cells (erythrocytes or kidney cells) with A blood group anti gen on their surface. High-avidity immune antibodies were found to be more efficient than 'natural' anti-A antibodies in this test. No evidence was found of anti-A antibodies being adsorbed on to the passing cells as tested by in vitro serological tests and tissue culture experiments. The applications of a technique for separating cells according to their surface antigens are con sidered obvious.