Antifluorescein affinity columns. Isolation and immunocompetence of lymphocytes that bind fluoresceinated antigens in vivo or in vitro.

Abstract

A new method for the isolation of specific immunocompetent [mouse or rat] lymphocytes was described in which lymphocyte populations are exposed to fluoresceinated antigens (FLAGs) in vivo or in vitro, and the FLAG-binding cells retained on antifluorescein affinity columns. Specific cells are then eluted with an unrelated FL-labeled protein and shown to be fully immunocompetent. This methodology was applied successfully in diverse antigenic systems including polymerized flagellin [Salmonella adelaide] and TNP [trinitrophenol] specific B [bone marrow derived] cells and alloantigen-reactive cytoxic T [thymus derived] lymphocytes. The method is rapid, inexpensive (requiring only antifluorescein beads), and can be applied to any antigens (or antibodies) in which a fluorescein group can be introduced.