Monoclonal antibodies to rat brain astroglial cells- their characterization and application for isolation of astroglial subpopulations

Abstract

Three monoclonal antibodies (MAbs) specifically recognizing rat astrocyte cell surface proteins have been characterized and their antigen binding specificities determined. One of these MAbs has been employed to isolate a distinct subpopulation of astroglial cells using immunoaffinity chromatography. MAbs to rat astroglial cell surface proteins, generated by fusion of mouse Sp2/O-Ag 14 myeloma cells and spleen cells from Balb/C mice immunized with purified astroglial cells, were screened for their cell binding specificities using ELISA and indirect immunofluorescence assay. The antigen binding specificity of three of these clones, which displayed specific binding to astrocytes, was determined by radioiodination of whole astrocytes and precipitation of the iodinated surface proteins by the MAbs. Immunoaffinity chromatography, using IgG from one of the clones coupled to CNBr activated Sepharose 6MB, demonstrated the potential usefulness of such MAbs in isolating a specific subpopulation of astroglial cells.