Detection of anti-p53 antibodies by ELISA using p53 synthetic or phage-displayed peptides.

Anti-p53 antibodies have been detected in the sera of patients with various types of cancers. In this report, we describe the development of a new ELISA aimed at detecting anti-p53 antibodies using two peptides belonging to immunodominant epitopes of the p53 N-terminal region. We first tested the reactivity of the sera by an indirect ELISA using the peptides as a capture system. Then, the specificity of the reaction was confirmed by an inhibition assay. Two systems of peptide presentation, phage display and the streptavidin/biotin system, were evaluated. Using a panel of sera from cancer patients, both systems were found to be equally reliable, demonstrating that both peptide-based ELISAs can be used for the specific detection of anti-p53 antibodies. The presence of anti-p53 antibodies was associated with p53 alteration whether it be mutation or accumulation.

[1]  S. H. van der Burg,et al.  Antibodies against p53 are associated with poor prognosis of colorectal cancer. , 1995, British Journal of Cancer.

[2]  B. Vogelstein,et al.  p53 mutations in human cancers. , 1991, Science.

[3]  T. Soussi,et al.  Linear antigenic sites defined by the B-cell response to human p53 are localized predominantly in the amino and carboxy-termini of the protein. , 1994, Oncogene.

[4]  M. Arima,et al.  Detection of serum p53 antibodies in patients with esophageal squamous cell carcinoma: correlation with clinicopathologic features and tumor markers. , 1998, Oncology reports.

[5]  J. Bourhis,et al.  Analysis of p53 serum antibodies in patients with head and neck squamous cell carcinoma. , 1996, Journal of the National Cancer Institute.

[6]  H. Schägger,et al.  Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. , 1987, Analytical biochemistry.

[7]  D. Pim,et al.  Detection of antibodies against the cellular protein p53 in sera from patients with breast cancer , 1982, International journal of cancer.

[8]  T. Soussi,et al.  Serum p53 antibodies as early markers of lung cancer , 1995, Nature Medicine.

[9]  C. Harris,et al.  Anti-p53 antibodies in sera of workers occupationally exposed to vinyl chloride. , 1995, Journal of the National Cancer Institute.

[10]  P. Laurent-Puig,et al.  Analysis of p53 antibodies in patients with various cancers define B-cell epitopes of human p53: distribution on primary structure and exposure on protein surface. , 1993, Cancer research.

[11]  E. Diamandis,et al.  Prevalence of serum antibodies against the p53 tumor suppressor gene protein in various cancers , 1994, International journal of cancer.

[12]  D. Laune,et al.  Improved performances of spot multiple peptide synthesis. , 1996, Peptide research.

[13]  T. Soussi,et al.  Production of human p53 specific monoclonal antibodies and their use in immunohistochemical studies of tumor cells. , 1993, Bulletin du cancer.

[14]  J. Minna,et al.  Antibodies against autologous tumor cell proteins in patients with small-cell lung cancer: association with improved survival. , 1993, Journal of the National Cancer Institute.

[15]  M. Hollstein,et al.  Circulating anti-p53 antibodies in esophageal cancer patients are found predominantly in individuals with p53 core domain mutations in their tumors. , 1996, Cancer research.

[16]  B. Milleron,et al.  Monitoring of p53 autoantibodies in lung cancer during therapy: relationship to response to treatment. , 1998, Clinical cancer research : an official journal of the American Association for Cancer Research.

[17]  A. Levine,et al.  The p53 tumour suppressor gene , 1991, Nature.

[18]  E. Appella,et al.  Structural mimicry and enhanced immunogenicity of peptide epitopes displayed on filamentous bacteriophage. The V3 loop of HIV-1 gp120. , 1994, Journal of molecular biology.

[19]  E. Diamandis,et al.  Humoral immune response against p53 protein in patients with colorectal carcinoma , 1997, International journal of cancer.

[20]  T. Soussi,et al.  p53 antibodies in the sera of lung cancer patients: Comparison with p53 mutation in the tumour tissue , 1995, International journal of cancer.

[21]  H Stein,et al.  Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). , 1984, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[22]  J. Bonneterre,et al.  Prognostic significance of circulating P53 antibodies in patients undergoing surgery for locoregional breast cancer , 1995, The Lancet.

[23]  R. Perham,et al.  New vectors for peptide display on the surface of filamentous bacteriophage. , 1996, Gene.

[24]  M. Hollstein,et al.  Anti-p53 antibodies in patients with Barrett's esophagus or esophageal carcinoma can predate cancer diagnosis. , 1998, Gastroenterology.

[25]  J. Kouyoumdjian,et al.  Detection and monitoring of serum p53 antibodies in patients with colorectal cancer. , 1997, Gut.

[26]  R. Perham,et al.  Simultaneous display of different peptides on the surface of filamentous bacteriophage. , 1997, Nucleic acids research.

[27]  K. Chandrasekaran,et al.  Presence of circulating antibodies against cellular protein p53 in a notable proportion of children with B‐cell lymphoma , 1987, International journal of cancer.

[28]  R. Sobti,et al.  A study on p53 protein and anti-p53 antibodies in the sera of patients with oesophageal cancer. , 1998, Mutation research.

[29]  T. Soussi,et al.  Analyses of p53 antibodies in sera of patients with lung carcinoma define immunodominant regions in the p53 protein. , 1994, British Journal of Cancer.

[30]  D. Marvin,et al.  Role of capsid structure and membrane protein processing in determining the size and copy number of peptides displayed on the major coat protein of filamentous bacteriophage. , 1996, Journal of molecular biology.

[31]  J. Jett,et al.  Anti-p53 antibodies in sera from patients with chronic obstructive pulmonary disease can predate a diagnosis of cancer. , 1996, Clinical cancer research : an official journal of the American Association for Cancer Research.

[32]  R. Perng,et al.  Presence of serum anti-p53 antibodies is associated with pleural effusion and poor prognosis in lung cancer patients. , 1998, Clinical cancer research : an official journal of the American Association for Cancer Research.

[33]  D. Lane,et al.  The immune response to p53 in breast cancer patients is directed against immunodominant epitopes unrelated to the mutational hot spot. , 1992, Cancer research.

[34]  Lubin,et al.  p53 antibodies in patients with various types of cancer: assay, identification, and characterization. , 1995, Clinical cancer research : an official journal of the American Association for Cancer Research.

[35]  D. Thomson,et al.  Analysis of the anti-p53 antibody response in cancer patients. , 1993, Cancer research.

[36]  J. Morrissey,et al.  Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. , 1981, Analytical biochemistry.

[37]  A. Larsson,et al.  P53 auto-antibodies in non-small cell lung cancer patients can predict increased life expectancy after radiotherapy. , 1998, Anticancer research.

[38]  T. Soussi,et al.  Critical residues of epitopes recognized by several anti-p53 monoclonal antibodies correspond to key residues of p53 involved in interactions with the mdm2 protein. , 2000, Journal of immunological methods.