Separation of cells by affinity chromatography on SpA-sepharose 6MB

A cell separation technique was designed which was based on the reaction of protein A of Staphylococcus aureus (SpA) with the Fc regions of IgG antibodies. SpA covalently linked to Sepharose 6MB is able to fix antibody-bearing cells, while non-fixed cells are removed by washing. The fixed cells can be released by dog IgG or by mechanical treatment. As an example of the use of the technique, Ig-bearing cells were isolated from mouse spleen cells treated with rabbit IgG anti-mouse Ig antibody; the purity of the Ig-bearing lymphocytes so isolated was better than 90%. The viability and the ability of the cells to shed the antibodies from their surface were not significantly impaired by the fractionation method. The technique can be generally applied to cell separation providing IgG antibodies against specific surface markers are available.