Development of ELISA method for detecting serum p53 antibodies

Objective To develop a specific indirect ELISA method for detecting anti-p53 antibodies in serum.Methods The recombinant p53 protein was used as the coating antigen to test sensitivity,specificity,precision and stability of ELISA method.203 serum samples from healthy individuals and 548 from patients with malign tumor were examined.Results The optimal concentration of the recombinant protein was(1 μg/mL) and the coating buffer was(0.05 mol/L) carbonic acid buffer(pH9.5).3 sera remained positive when they were diluted by the ration of(1∶400),and the variation coefficient was 3.67%-16.88% after repeatedly assaying for 8 times.Blocking test of specific IgG showed the rate of blocking increased with the p53 protein.Serum samples from healthy persons observed no positive(0/203) whereas 14.23%(78/548) of patients with tumor were positive. Conclusion The new developed ELISA method for p53 antibodies detection was wide range,fine sensitivity,high precision and stability.