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

J Med Virol. 1993 May;40(1):14-21. doi: 10.1002/jmv.1890400105.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Viral / blood*
  • Baculoviridae / genetics
  • Base Sequence
  • Blotting, Western
  • Epitopes / chemistry
  • Epitopes / genetics
  • Epitopes / immunology
  • False Negative Reactions
  • Female
  • Humans
  • Molecular Sequence Data
  • Oncogene Proteins, Viral / chemistry
  • Oncogene Proteins, Viral / genetics
  • Oncogene Proteins, Viral / immunology*
  • Open Reading Frames
  • Papillomaviridae / genetics
  • Papillomaviridae / immunology*
  • Papillomaviridae / isolation & purification
  • Papillomavirus E7 Proteins
  • Radioimmunoprecipitation Assay
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Tumor Virus Infections / immunology
  • Tumor Virus Infections / microbiology*
  • Uterine Cervical Neoplasms / immunology
  • Uterine Cervical Neoplasms / microbiology*

Substances

  • Antibodies, Viral
  • Epitopes
  • Oncogene Proteins, Viral
  • Papillomavirus E7 Proteins
  • Recombinant Proteins
  • oncogene protein E7, Human papillomavirus type 16