展示p53蛋白表位噬菌体phage-SD的制备及应用  

Preparation and application of bacteriophage phage-SD displaying epitope of p53 protein

作　　者：刘兴友 孙梦月[1] 潘鹏涛 刘国伟 Liu Xingyou;Sun Mengyue;Pan Pengtao;Liu Guowei(College of life sciences,Henan Normal University,Xinxiang 453007,China;College of Life Science and Basic Medicine,Xinxiang University,Xinxiang 453003,China;Xinxiang Central Hospital,Xinxiang 453099,China)

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]新乡学院生命科学与基础医学学院,河南新乡453003 [3]新乡市中心医院,河南新乡453099

出　　处：《河南师范大学学报（自然科学版）》2023年第2期109-114,共6页Journal of Henan Normal University(Natural Science Edition)

基　　金：国家自然科学基金(81702074);河南省科技攻关计划项目(212102110160);河南省高等学校重点科研项目(22A180008).

摘　　要：[目的]利用噬菌体展示技术制备一种可用于检测血清p53抗体的噬菌体.[方法]利用基因工程手段将编码p53蛋白N端20~31位的多肽SD的基因克隆到丝状噬菌体载体fADL-le的pIII蛋白基因中,制备展示该外源肽的噬菌体phage-SD并进行了Western blot鉴定.在此基础上,分别以phage-SD和重组p53蛋白为检测抗原,利用ELISA方法对乳腺癌患者血清p53抗体进行检测.[结果]成功制备出能特异性识别血清p53抗体的噬菌体phage-SD.ELISA检测结果表明,60例乳腺癌患者中phage-SD检测到13例p53抗体阳性患者,检出率为21.67%,与重组p53蛋白检测效率(21.67%)一致.[结论]成功制备一种灵敏度高、特异性强的可用于血清p53抗体检测的噬菌体phage-SD,为新型血清p53抗体检测试剂的开发及临床应用奠定基础.[Objective]This research aimed to prepare a new type of phage using phage display technology for the detection of serum p53 antibodies.[Methods]The fADL-le phage vector was double digested by Bgl I and then the adaptor molecule encoding the peptide SD located in the N-terminal of 20-31 amino acid of p53 protein was inserted in the gene III,and phage-SD was prepared and analyzed by Western blot.Then,serum p53 antibody in the breast cancer patients was detected by ELISA method using phage-SD and recombinant p53 protein as the coating antigen,respectively.[Results]The peptide SD was successfully displayed on the surface of the bacteriophage and the expressed peptide SD also could identify the p53 antibody.Among the 60 breast cancer patients,17 patients(21.67%)were tested positive for serum p53 antibody by phage-SD ELISA,and the detection efficiency of this method was the same as the p53-ELISA.[Conclusion]Phage-SD was successfully prepared,and this phage presented high detection efficiency and specificity and had the potential to develop into a new type of diagnosis reagent for the detection of serum p53 antibody.

关 键 词：噬菌体展示 丝状噬菌体 P53抗体 ELISA 

分 类 号：R-33[医药卫生]