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 unc hanged 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 demons trated, whereas IgG2, IgG4 and IgE antibodies did not bind. Thus, cova lently coupled Penicillin-G can be used to study the immune-response t o the unchanged beta-lactam ring in patients receiving penicillin ther apy. The new method is complementary to hitherto described techniques, which generally only allow detection of antibodies binding to penicil loyl-groups.