Cell sorting using a universally applicable affinity chromatography matrix: solid-phase anti-fluorescein isothiocyanate antibody.
Procedures are described for fractionating cells utilizing a universally applicable cellular affinity chromatography matrix. The affinity matrix consists of immunoabsorption purified goat anti-fluorescein isothiocyanate antibody coupled to large derivatized polyacrylamide beads. This matrix may, in principle, be used to isolate any cell subpopulation provided it has a fluorescein-labeled ligand on its surface. In this report the matrix was used to isolate viable purified fractions of mouse surface Ig-positive cells, Lyt1 cells, and mouse lymphocytes that bind the lectin soybean agglutinin. A preliminary experiment using the anti-FITC beads suggested that this technique can provide a fraction of cells enriched in antigen binding cells. Cell populations isolated by this technique retain their ability to respond to in vitro mitogen stimulation, as well as their ability to be maintained in cell culture following fractionation. Additional experiments using a column consisting of goat anti-rabbit Ig antibody coupled to the same support material are also reported.
D000818:Animals; D000906:Antibodies; D002469:Cell Separation / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods*; D005260:Female; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins / Q000276:immunology*; D007152:Immunologic Capping; D008213:Lymphocyte Activation; D051379:Mice; D008813:Mice, Inbred ICR; D008815:Mice, Inbred Strains; D011947:Receptors, Antigen, B-Cell / Q000302:isolation & purification; D013861:Thiocyanates / Q000276:immunology*
D000906:Antibodies; D005452:Fluoresceins; D011947:Receptors, Antigen, B-Cell; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate