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 lymphocytes has been 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 has been applied successfully in diverse antigenic systems including polymerized flagellin and TNP-specific B cells and alloantigen-reactive cytotoxic T 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.

Memory B cells from a subset of treatment‐naïve relapsing‐remitting multiple sclerosis patients elicit CD4+ T‐cell proliferation and IFN‐γ production in response to myelin basic protein and myelin oligodendrocyte glycoprotein

“…In total, 0.1% NaN 3 was added to the conjugated protein preparations as preservative. OVA‐FITC was generated to detect background of OVA‐ (and FITC)‐reactive B cells 68.…”

Genetics, Cellular Expression and Function of IgD and IgM Receptors

“…An antiserum with similar specificity was generated by Ahmed et al (1977), who absorbed a C57BL/6 anti-DBA/2 antiserum * spleen cells from 3-week-old mice were treated with anti-5 alloantisemm, washed, and labeled with fluorescein-conjugated goat anti-y serum. ** The labeled cells were then passed over an anti-fiuorescein aflRnity column as described (Scott 1976). The percentage of IgD-positive C57BIV6.Ig« cells in the effluent ('passed') was reduced by about 70%.…”

B‐Lymphocyte Alloantigens

“…subpopulations enriched for B cells or for T cells were prepared by the recently developed method of anti-fluorescein affinity column chromotography (11). Affinity-purified pony anti-fluorescein keyhole limpet hemocyanin (KLH) was conjugated to Sepharose 4B.…”