噬菌体随机12肽库筛选肝癌患者血清肿瘤标记物的初步研究

Screening of phage random peptide library with sera from patients with liver cancer

目的:从噬菌体随机多肽文库中,筛选出能与肝癌患者血清特异性结合的短肽分子。方法:采用肝癌患者血清作为配基,筛选以融合蛋白形式在丝状噬菌体M13外壳蛋白III表达的随机12肽文库。按吸附-洗脱-扩增的淘筛过程,经3轮淘筛后,随机挑取噬菌体克隆用ELISA检测其特异性,评价分析其诊断肝癌的价值。结果:经3轮淘筛后,特异性结合的噬菌体富集增加近100倍。用ELISA检测第3轮筛选后随机挑取的单个噬菌体克隆,其中特异性最好的3个克隆具有诊断肝癌的潜在价值。结论:噬菌体展示肽库技术,可以有效进行肝癌相关抗原肽的筛选研究,为获得特异性诊断试剂进而为肝癌的诊断提供依据。

Objective：To obtain the short peptides through screening phage random peptide library with sera from liver cancer patients Methods： Sera of patients with liver cancer was used to immunoscreen a phage random peptide library of 12 anmino - acid residues displayed as a fusion protein n shell protein III of filamentous phage M13. The positive clones were obtained through three rounds of biopanning;the reactivity of each clone bound to the sera was examined by ELISA. Results： The eluted phages were enriched nearly to 100 fold through three rounds of biopanning. From the clones selected randomly from the third round biopanning, we selected the best 3 clones which could bind to the sera sera from liver cancer patients. Conclusion： Liver cancer - specific peptides can be identified by the technique of immunoscreening phage random peptide library using the patients sera and these peptides pay the foundation for preparing immunodiagnostic reagent or vaccine.