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 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.
Wlazlo
A P
AP
The Wistar Institute, Philadelphia, PA 19104, USA.
Giles-Davis
W
W
Clements
A
A
Struble
G
G
Marmorstein
R
R
Ertl
H C
HC
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
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