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.
Publication types
-
Research Support, Non-U.S. Gov't
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
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
Substances
-
Antibodies, Monoclonal
-
Antibodies, Viral
-
Antigens, Viral
-
DNA Primers
-
E6 protein, Human papillomavirus type 16
-
Oncogene Proteins, Viral
-
Papillomavirus E7 Proteins
-
Peptide Fragments
-
Repressor Proteins
-
oncogene protein E7, Human papillomavirus type 16