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Background: Although clinical significance of serum p53 Abs (s-p53 Abs) in cancer patients has been studied during the past few years, how to detect it effectively is a crucial issue. Methods: wild-type p53 protein (wt p53) and phage-displayed peptide were prepared. Three types of enzymelinked immunosorbent assays (ELISA) methods were constructed and a hybrid antigen-ELISA approach was attempted. Results: 7 breast cancer patients (19.4%) were detected s-p53 Abs positive by p53-ELISA, 4 patients (11.1%) were detected positive by phage-ELISA, and 10 patients (27.8%) were detected positive by p53-phage ELISA. A combination of 1.5μg/ml wt p53 and 40μg/ml phage was the best concentration for a mix antigen in hybrid antigen-ELISA. Conclusion: A combination of p53-ELISA and phage-ELISA can effectively improve the sensitivity of s-p53 Abs. This increases the application possibility of s-p53 Abs on clinic.