Laboratory of Biomass Valorisation and Production of Eukaryotic Proteins, Centre of Biotechnology of Sfax, University of Sfax, Route Sidi Mansour, BP 1177, 3038 Sfax, Tunisia.
Mol Biol Rep. 2013 May;40(5):3865-72. doi: 10.1007/s11033-012-2467-1. Epub 2013 Mar 26.
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.
血清方法检测 P53 改变更容易进行,不需要肿瘤组织,并且对患者监测具有重要意义。在这项研究中,我们描述了一种基于重组人 P53 蛋白的自制 ELISA 检测方法的开发,该蛋白由巴氏毕赤酵母生产,用作检测结直肠癌患者血清 p53-Abs 的抗原。人 P53 通过重组 KM71 菌株(Kα21)通过酿酒酵母交配型基因的肽信号α作为 His 标记蛋白进行分泌。重组 P53-His 能够在 23.4%的患者中检测到 p53-Abs。血清 p53-Abs 与手术治疗显著相关(P = 0.007),随访期间复发(P = 0.036),浸润深度(P = 0.036)和 CA19-9 水平(P = 0.034)。生存分析显示,血清 p53-Abs 阴性的患者无疾病生存时间延长(P log rank = 0.012)。总之,巴氏毕赤酵母分泌的重组人 P53-His 似乎是检测结直肠癌患者血清 p53 Abs 的有用抗原。