Hapten-protein conjugates prepared by the mixed anhydride method. Cross-reactive antibodies in heterologous antisera

Poly-L-lysine conjugates of three structurally unrelated mycotoxins were made by either a mixed anhydride intermediate (MA) or an activated N-hydroxysuccinimide ester intermediate (AE). Control conjugates, with no mycotoxin, were also prepared by each method. Antisera elicited by mycotoxin-albumin conjugates produced by the MA method bound to the three poly-L-lysine-MA mycotoxin conjugates and the MA control conjugate , but bound only to the poly-L-lysine-AE conjugates of the homologous mycotoxin. Binding of antisera to homologous poly-L-lysine conjugates was always inhibited by free hapten when the conjugate was prepared by the AE method but not by the MA method. The specific inhibition of antibody binding by various synthetic haptens indicated that the cross-reactions associated with the MA method were due to the undesired conjugation of isobutylformate during the mixed anhydride procedure.