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Mol Biol Rep ; 40(5): 3865-72, 2013 May.
Article in English | MEDLINE | ID: mdl-23526366

ABSTRACT

The detection of P53 alteration by serological method is easier to perform, does not require tumor tissues and is of interest for patients monitoring. In this study, we described the development of a home made ELISA test based on recombinant human P53 protein produced in Pichia pastoris and used as antigen for the detection of serum p53-Abs in colorectal carcinoma patients. The human P53 was secreted as a His-tagged protein by recombinant KM71 strain (Kα21) via the peptide signal α of the Saccharomyces cerevisiae mating type gene. The recombinant P53-His was able to detect p53-Abs in 23.4% of patients. Serum p53-Abs correlated significantly with surgical treatment (P = 0.007), relapse during follow-up (P = 0.036), depth of invasion (P = 0.036) and the level of CA19-9 (P = 0.034). Survival analysis showed that patients negative for serum p53-Abs exhibited a prolonged disease free survival period (P log rank = 0.012). In conclusion, the secreted recombinant human P53-His produced in P. pastoris seems to be a useful antigen for detection of serum p53 Abs in patients with colorectal carcinoma.


Subject(s)
Adenocarcinoma/diagnosis , Adenocarcinoma/immunology , Autoantibodies/blood , Colorectal Neoplasms/diagnosis , Colorectal Neoplasms/immunology , Pichia/genetics , Tumor Suppressor Protein p53/immunology , Adenocarcinoma/blood , Adenocarcinoma/mortality , Adult , Aged , Antibodies, Neoplasm/blood , Antibodies, Neoplasm/immunology , Antigens, Neoplasm/blood , Antigens, Neoplasm/immunology , Colorectal Neoplasms/blood , Colorectal Neoplasms/mortality , Female , Humans , Male , Middle Aged , Neoplasm Staging , Pichia/metabolism , Recombinant Fusion Proteins/blood , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/isolation & purification , Tumor Suppressor Protein p53/blood , Tumor Suppressor Protein p53/genetics
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