A highly sensitive enzyme-linked immunosorbent assay for idiotype-bearing antibodies.
A sensitive and specific immunoenzyme assay (ELISA) for quantitation of total and cross-reactive idiotype-bearing (CRI) anti-ABA antibodies is described. Total anti-ABA antibodies are directly assessed in ABA-BGG coated polyvinyl wells with enzyme-labelled rabbit anti-mouse immunoglobulins. By interpolation on a standard curve absorbance values give the concentration of anti-ABA antibodies with a sensitivity of 30 ng/ml. CRI+ antibodies are quantitated by inhibition of enzyme-labelled monoclonal CRI+ antibody binding to solid-phase coated rabbit anti-CRI immunoglobulins. The concentration of CRI+ antibodies, evaluated by interpolation on a standard inhibition curve, can be measured at the level of 10 ng/ml. This highly sensitive, rapid, specific and reproducible assay is easily used, with minor modifications, to detect specific antibodies in any idiotype system.
D000818:Animals; D000918:Antibody Specificity; D001391:Azo Compounds / Q000276:immunology*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D007124:Immunoenzyme Techniques; D007130:Immunoglobulin Idiotypes / Q000032:analysis* / Q000378:metabolism; D051379:Mice; D008805:Mice, Inbred A; D011817:Rabbits; D010132:p-Azobenzenearsonate / Q000276:immunology*
D001391:Azo Compounds; D007130:Immunoglobulin Idiotypes; D010132:p-Azobenzenearsonate