Generation and characterization of monoclonal antibodies against the E6 and E7 oncoproteins of HPV

Generation of three monoclonal antibodies (MAbs) to the major oncoproteins of human papillomavirus (HPV) was accomplished by an intense prime/boost re gimen. Mice were primed with expression vectors expressing either the E6 or E7 oncoproteins of HPV-16 followed by boosting with a vaccinia virus const ruct and a replication-defective E1-deleted adenoviral recombinant of the h uman strain 5, and last, with baculovirus-derived HPV-16 E6 and E7 proteins in incomplete Freunds' adjuvant. Splenocytes were then fused with a myelom a cell line. The vaccination protocol generated one anti-E7 MAb of the IgM isotype and two anti-E6 MAbs of the IgG1 subisotype. The MAbs were tested f or functionality in standard laboratory assays and found to detect the E6 a nd E7 proteins, respectively. The E7 MAb cross-reacted with the HPV-1a E7 o ncoprotein. The binding sites of the MAbs were mapped to defined regions of each viral protein.