A solid phase immunofluorometric assay for anti-lymphocyte antibodies.
A solid phase immunofluorometric assay for anti-lymphocyte antibodies was developed by using agarose beads to which partially purified HLA antigens were bound. Optimal conditions for the assay system was determined with respect to bead concentration, serum concentration, FTC-conjugate concentration and incubation times. This system detected non-complement fixing as well as complement fixing antibodies in antisera. It was approximately 20 times more sensitive than the microlymphocytotoxicity test or an indirect immunofluorescence test in which viable cells were used. The antigen bound to the bead was stable at least for 54 days at 5 degrees C or - 80 degrees C. When lyophilized, its antibody-binding activity was 80% of the original. By this system, pregnancy sera containing lymphocytotoxic antibodies and normal sera gave relative fluorescence intensities of 331 +/- 65 (X +/- S.D.; n = 8) and 188 +/- 54 (n = 9), respectively.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D000941:Antigens; D000961:Antilymphocyte Serum / Q000032:analysis*; D001666:Binding Sites, Antibody; D003168:Complement Fixation Tests; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D007074:Immunoglobulin G; D007518:Isoantibodies; D008297:Male; D011247:Pregnancy; D011817:Rabbits; D013205:Staphylococcal Protein A / Q000494:pharmacology; D013997:Time Factors
D000888:Antibodies, Anti-Idiotypic; D000941:Antigens; D000961:Antilymphocyte Serum; D007106:Immune Sera; D007074:Immunoglobulin G; D007518:Isoantibodies; D013205:Staphylococcal Protein A