Monoclonal antibodies to a benzo[a]pyrene diolepoxide modified protein

Abstract

Monoclonal antibodies have been developed which specifically recognize protein modified by 7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE-I). These antibodies have been characterized as to sensitivity and specificity by an enzyme-linked immunosorbent assay. They react with BPDE-I-modified proteins and do not cross-react with non-modified protein. All the antibodies have higher sensitivity toward BPDE-I-tetraols than to BPDE-I-modified protein. They cross-react with BPDE-I-deoxyguanosine, BPDE-I-DNA and BPDE-II-rabbit serum albumin. Because of the high antibody sensitivity for tetraols, an immunoassay was developed for quantitating DNA and protein adducts based on release of tetraols from modified samples. This approach was validated using radiolabeled BPDE-I modified DNA and protein.