INDUCTION AND CHARACTERIZATION OF MULTIANALYTE ANTIBODIES AGAINST PENICILLINS IN EGG-YOLK

Antibodies against penicillins were induced in eggs of laying hens aft er immunization with 6-aminopenicillanic acid (6-APA) coupled to keyho le limpet hemocyanin (KLH). Development of the antibody titer was moni tored by an indirect enzyme-linked immunosorbent assay (ELISA), with 6 -APA coupled to ovalbumin as antigen for coating microtiter plates. Di fferent characteristics (time course, affinity) of the immune reaction were observed by testing eggs of individual hens. Titer values varied between 150 and 2000. Antibodies were isolated by polyethylene glycol precipitation and affinity chromatography, using a hapten sorbent wit h 6-APA as ligand, Glycine buffer, pH 3.0, was used to elute the immun oglobulins. Antibody specificity was determined in a competitive ELISA with 7 penicillins and the cephalosporin cephalexin as competitors. C ross reactivities for the penicillins were between 100 and 290% (6-APA = 100%). Cephalexin cross reacted only marginally (3%).