title: Generation and characterization of monoclonal antibodies against the E6 and E7 oncoproteins of HPV.
abstract: Generation of three monoclonal antibodies (MAbs) to the major oncoproteins of human papillomavirus (HPV) was accomplished by an intense prime/boost regimen. Mice were primed with expression vectors expressing either the E6 or E7 oncoproteins of HPV-16 followed by boosting with a vaccinia virus construct and a replication-defective E1-deleted adenoviral recombinant of the human strain 5, and last, with baculovirus-derived HPV-16 E6 and E7 proteins in incomplete Freunds' adjuvant. Splenocytes were then fused with a myeloma 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 for functionality in standard laboratory assays and found to detect the E6 and E7 proteins, respectively. The E7 MAb cross-reacted with the HPV-1a E7 oncoprotein. The binding sites of the MAbs were mapped to defined regions of each viral protein.
authors.affiliation: The Wistar Institute, Philadelphia, PA 19104, USA.
authors.forename: A P
authors.initials: AP
authors.lastname: Wlazlo
authors.forename: W
authors.initials: W
authors.lastname: Giles-Davis
authors.forename: A
authors.initials: A
authors.lastname: Clements
authors.forename: G
authors.initials: G
authors.lastname: Struble
authors.forename: R
authors.initials: R
authors.lastname: Marmorstein
authors.forename: H C
authors.initials: HC
authors.lastname: Ertl
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis*; D000914:Antibodies, Viral / Q000032:analysis / Q000096:biosynthesis*; D000917:Antibody Formation; D000956:Antigens, Viral / Q000276:immunology*; D015153:Blotting, Western; D002478:Cells, Cultured; D017931:DNA Primers / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000276:immunology* / Q000302:isolation & purification; D027383:Papillomaviridae / Q000276:immunology*; D050725:Papillomavirus E7 Proteins; D010446:Peptide Fragments / Q000276:immunology; D016133:Polymerase Chain Reaction; D012097:Repressor Proteins; D017386:Sequence Homology, Amino Acid
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D017931:DNA Primers; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D010446:Peptide Fragments; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16