Content area
Full Text
Mol Biol Rep (2013) 40:38653872 DOI 10.1007/s11033-012-2467-1
Secreted recombinant P53 protein from Pichia pastoris is a useful antigen for detection of serum p53: autoantibody in patients with advanced colorectal adenocarcinoma
Salma Abdelmoula-Souissi Nourane Zouari Imen Miladi-Abdenadher
Ouhoud Yaich-Kolsi Ines Ayadi-Masmoudi Abdelmajid Khabir
Hatem Masmoudi Mounir Frikha Raja Mokdad-Gargouri
Received: 23 May 2012 / Accepted: 18 December 2012 / Published online: 26 March 2013 Springer Science+Business Media Dordrecht 2013
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 (Ka21) via the peptide signal a 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 signicantly 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.
Keywords Anti-p53 antibody Pichia pastoris
Secreted recombinant p53 protein Colorectal cancer
Introduction
The tumour suppressor P53 is a key player to maintain genomic stability in response to DNA damage, hypoxia and oncogene activation [1, 2]. The main mechanism by which P53 ensures its protective roles is as a transcription factor. Indeed, by binding to specic DNA sequence, P53 modulates the transcription of key genes involved in cell cycle arrest, apoptosis, DNA repair and inhibition of neo-angiogenesis [3]. Inactivation of the p53 gene is the most common genetic alteration observed in human cancers [2]. Indeed about 50 % of tumors have a mutated P53 which escape to mdm2 mediated proteasome degradation and became more stable with a prolonged half-life [4]. Thereby, the accumulated P53 protein are recognized as...