ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR DETECTION OF ANTIBODIES TO THE UNMODIFIED BETA-LACTAM RING

Dose- and pH- dependent carbodiimide-mediated coupling of Penicillin-G to polystyrene microtiter-plates that leaves the beta-lactam ring unchanged is described. A new ELISA method was developed using Penicillin-G coated plates. The binding of 3 different monoclonal antibodies as well as human IgG antibodies of the IgG, and IgG3 subclasses is demonstrated, whereas IgG2, IgG4 and IgE antibodies did not bind. Thus, covalently coupled Penicillin-G can be used to study the immune-response to the unchanged beta-lactam ring in patients receiving penicillin therapy. The new method is complementary to hitherto described techniques, which generally only allow detection of antibodies binding to penicilloyl-groups.