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Clinical Chemistry 41: 1720-1729, 1995;
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Clinical Chemistry, Vol 41, 1720-1729, Copyright © 1995 by American Association for Clinical Chemistry

Time-resolved immunofluorometric assay of p53 protein

MA Levesque, M D'Costa, K Angelopoulou and EP Diamandis
Department of Pathology, Mount Sinai Hospital, Toronto, Ontario, Canada.

A common feature of human tumor tissue is mutant p53 protein accumulation. Here we evaluate a new "sandwich" immunoassay for p53 protein incorporating modifications to a previously reported method, including the use of microtiter plates coated directly with the anti- p53 monoclonal antibody DO-1, a detergent- and mouse serum-containing sample diluent, and a labeled secondary antibody diluent containing goat serum. The use of CM-1 antiserum to probe the immunocaptured p53 and the detection of bound complexes by a labeled secondary antibody allows coupling to a time-resolved fluorescence detection system. The new assay yielded p53 concentrations comparable with those by the previous assay for breast tumor cytosols (n = 198), nondiseased breast tissues (n = 70), and five transformed cell lines, but showed differences in p53 values measured in sera from patients without cancer (n = 78). These serum differences were found to reflect nonspecific interferences affecting the original method, which implies that the new immunoassay has improved specificity for serum p53 quantification.


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 Clin. Cancer Res.Home page
M. A. Levesque, D. Katsaros, M. Massobrio, F. Genta, H. Yu, G. Richiardi, S. Fracchioli, A. Durando, R. Arisio, and E. P. Diamandis
Evidence for a Dose-Response Effect between p53 (but not p21WAF1/Cip1) Protein Concentrations, Survival, and Responsiveness in Patients with Epithelial Ovarian Cancer Treated with Platinum-based Chemotherapy
Clin. Cancer Res., August 1, 2000; 6(8): 3260 - 3270.
[Abstract] [Full Text]


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