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.
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.
Stacey
S N
SN
Department of Molecular Biology, Paterson Institute for Cancer Research, Christie Hospital, Manchester, England.
Ghosh
A
A
Bartholomew
J S
JS
Tindle
R W
RW
Stern
P L
PL
Mackett
M
M
Arrand
J R
JR
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*
D000914:Antibodies, Viral; D000939:Epitopes; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D011994:Recombinant Proteins; C059731:oncogene protein E7, Human papillomavirus type 16