title: Expression of human papillomavirus type 16 E7 protein by recombinant baculovirus and use for the detection of E7 antibodies in sera from cervical carcinoma patients.
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.
authors.affiliation: Department of Molecular Biology, Paterson Institute for Cancer Research, Christie Hospital, Manchester, England.
authors.forename: S N
authors.initials: SN
authors.lastname: Stacey
authors.forename: A
authors.initials: A
authors.lastname: Ghosh
authors.forename: J S
authors.initials: JS
authors.lastname: Bartholomew
authors.forename: R W
authors.initials: RW
authors.lastname: Tindle
authors.forename: P L
authors.initials: PL
authors.lastname: Stern
authors.forename: M
authors.initials: M
authors.lastname: Mackett
authors.forename: J R
authors.initials: JR
authors.lastname: Arrand
mesh_terms: D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000097:blood*; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D005188:False Negative Reactions; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D016366:Open Reading Frames; D027383:Papillomaviridae / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D050725:Papillomavirus E7 Proteins; D015531:Radioimmunoprecipitation Assay; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014412:Tumor Virus Infections / Q000276:immunology / Q000382:microbiology*; D002583:Uterine Cervical Neoplasms / Q000276:immunology / Q000382:microbiology*
chemical_list: D000914:Antibodies, Viral; D000939:Epitopes; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D011994:Recombinant Proteins; C059731:oncogene protein E7, Human papillomavirus type 16