Separation of human lymphoid cells by affinity chromatography and cell surface labelling by hydroxyethyl methacrylate particles using monoclonal antibodies

J Chromatogr. 1986 Apr 11:376:401-8.

Abstract

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.

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Cell Separation / methods
  • Chromatography, Affinity
  • Humans
  • Immunoenzyme Techniques
  • Immunosorbents
  • Lymphocytes / cytology*
  • Methacrylates
  • Palatine Tonsil / cytology
  • Surface Properties
  • Swine
  • Thymidine / metabolism

Substances

  • Antibodies, Monoclonal
  • Immunosorbents
  • Methacrylates
  • hydroxyethyl methacrylate
  • Thymidine