Separation of human lymphoid cells by affinity chromatography and cell surface labelling by hydroxyethyl methacrylate particles using monoclonal antibodies.
The hydroxyalkyl methacrylate gel Separon, Sepharose 6MB and polystyrene dishes with bound antibodies were used for affinity chromatography of cells isolated from human tonsils and peripheral blood. Enrichment of T cells by filtration through anti-human immunoglobulin Separon or Sepharose 6MB columns was comparable to that achieved by "panning" on dishes or by Nylon wool column fractionation. High enrichment of B cells was obtained when cells pretreated with monoclonal anti-T cell antibodies were fractionated on columns or dishes where anti-mouse antibody was bound. Hydroxyethyl methacrylate copolymer microspheres were conjugated with monoclonal antibodies directed against surface markers of human lymphocytes or with anti-mouse immunoglobulin antibodies and used for surface labelling of cells. The results obtained by immunofluorescence were in good correlation with labelling by hydroxyethyl methacrylate immunobeads.
D000818:Animals; D000911:Antibodies, Monoclonal; D002469:Cell Separation / Q000379:methods; D002846:Chromatography, Affinity; D006801:Humans; D007124:Immunoenzyme Techniques; D007164:Immunosorbents; D008214:Lymphocytes / Q000166:cytology*; D008689:Methacrylates; D014066:Palatine Tonsil / Q000166:cytology; D013499:Surface Properties; D013552:Swine; D013936:Thymidine / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007164:Immunosorbents; D008689:Methacrylates; C005044:hydroxyethyl methacrylate; D013936:Thymidine