Abstract
Although the presence of serum antibodies against the human papillomavirus type 16 (HPV-16) E7 protein has been linked with cervical cancer, currently available assays detect antibodies in only ca. 40% of carcinoma patients. The dependence of these serological assays on synthetic target antigens which present only linear epitopes may be a limiting factor. In order to produce a more realistic target antigen for use in serological assays, we have expressed the HPV-16 E7 protein in insect cells using a recombinant baculovirus vector. Two major E7 forms of ca. 18kDa and 16kDa were produced and characterised. The 16kDa component was shown to be truncated at the N-terminus. A radioimmunoprecipitation assay was developed for the detection of anti-E7 antibodies in human sera. This assay showed a marked increase in detection rate compared with a western blotting method based on bacterially derived E7 fusion proteins.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Antibodies, Viral / blood*
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Baculoviridae / genetics
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Base Sequence
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Blotting, Western
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Epitopes / chemistry
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Epitopes / genetics
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Epitopes / immunology
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False Negative Reactions
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Female
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Humans
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Molecular Sequence Data
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Oncogene Proteins, Viral / chemistry
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Oncogene Proteins, Viral / genetics
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Oncogene Proteins, Viral / immunology*
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Open Reading Frames
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Papillomaviridae / genetics
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Papillomaviridae / immunology*
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Papillomaviridae / isolation & purification
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Papillomavirus E7 Proteins
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Radioimmunoprecipitation Assay
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
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Tumor Virus Infections / immunology
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Tumor Virus Infections / microbiology*
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Uterine Cervical Neoplasms / immunology
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Uterine Cervical Neoplasms / microbiology*
Substances
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Antibodies, Viral
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Epitopes
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Oncogene Proteins, Viral
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Papillomavirus E7 Proteins
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Recombinant Proteins
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oncogene protein E7, Human papillomavirus type 16