Demonstration of a 65 kDa tumor-specific phosphoprotein in urine and serum of rats with N-methyl-N-nitrosourea-induced mammary adenocarcinomas

摘要: Polyclonal antibodies against a 65 kDa tumor-associated phosphoprotein (p65) were used to develop an ELISA analyze the presence of p65 in urine and serum rats bearing N-methyl-N-nitrosourea-induced mammary gland adenocarcinomas. Highly purified rat was added normal establish quantitative standard curve with average correlation coefficient being 0.98 0.99 respectively. All samples obtained from different carcinoma-bearing showed concentrations above levels found control sera. The between tumor burden concentration 0.65 0.77 37.0 +/- 32.0 48.0 38.0 ng/ml In case adenocarcinomas, mean level 119.0 35.9 their 225.4 67.5 ng/ml. Sensitivity, specificity predictive value for marker elevation 78.5, 70.0 78.5% Following vitro phosphorylation concentrated urinary proteins, isoelectrofocusing, SDS-PAGE autoradiography, phosphorylated form protein pI 5.8 identified tumor-bearing rats. This bound antiphosphotyrosine monoclonal antibody anti-p65 polyclonal as determined by Western blot analysis. Using immunoprecipitation procedure, main radio- immunoactive band two lower mol. wt bands 50 41 kDa, apparently representing degradation products p65, after vivo proteins