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

• Published in: Hybridoma Vol. 20; no. 4; p. 257
• Main Authors: Wlazlo, A P, Giles-Davis, W, Clements, A, Struble, G, Marmorstein, R, Ertl, H C
• Format: Journal Article
• Language: English
• Published: United States 01.08.2001
• Subjects: Amino Acid Sequence; Animals; Antibodies, Monoclonal - analysis; Antibodies, Monoclonal - biosynthesis; Antibodies, Viral - analysis; Antibodies, Viral - biosynthesis; Antibody Formation; Antigens, Viral - immunology; Blotting, Western; Cells, Cultured; DNA Primers - chemistry; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Female; Humans; Mice; Mice, Inbred BALB C; Mice, SCID; Molecular Sequence Data; Oncogene Proteins, Viral - immunology; Oncogene Proteins, Viral - isolation & purification; Papillomaviridae - immunology; Papillomavirus E7 Proteins; Peptide Fragments - immunology; Polymerase Chain Reaction; Repressor Proteins; Sequence Homology, Amino Acid
• ISSN: 0272-457X
• DOI: 10.1089/027245701753179848

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.