人Rh(D)血型抗原模拟表位筛选及鉴定的临床意义  

Clinical significance of Screening and Identification of Human Rh (D) Blood Group Antigen Mimotopes

作　　者：张勤 ZHANG Qin(Laboratory Medicine Center,Zhengzhou Yihe Hospital,Zhengzhou 450047,China)

机构地区：[1]郑州颐和医院检验医学中心,河南郑州450047

出　　处：《医学临床研究》2021年第3期437-440,共4页Journal of Clinical Research

摘　　要：【目的】探讨人Rh(D)血型抗原模拟表位筛选及鉴定的临床意义。【方法】以抗Rh(D)单克隆抗体作为固相筛选分子,随机选取35个进行克隆,经噬菌体酶联免疫吸附测定法(ELISA)及交叉试验,确定阳性克隆,采用蛋白石免疫印迹法(Western blot)对噬菌体行抗原性分析。【结果】DNA序列测序及竞争抑制实验在11个克隆的噬菌体所展示的外源多肽中有4个克隆无共性,可能是非特异性结合。7个具有克隆氨基酸序列均含有相同的脯氨酸(P)、色氨酸(W)、谷氨酸(Q)结构(-WP-Q-),超过40%的抑制率。Western blot分析表明,被选定的噬菌体能被抗Rh(D)血清特异识别,抗原性与Rh(D)蛋白相类似。【结论】利用抗Rh(D)单克隆抗体筛选噬菌体随机肽库,可获得具有(-WP-Q-)结构的Rh(D)抗原模拟表位。【Objective】To investigate the clinical significance of screening and identification of mimic epitopes of human Rh(d)blood group antigen.【Methods】Anti Rh(d)monoclonal antibodies were used as solid phase screening molecules,and 35 of them were randomly selected for cloning.The positive clones were identified by phage enzyme-linked immunosorbent assay(ELISA)and cross test.The antigenicity of phage was analyzed by Western blot.【Results】The results of DNA sequencing and competitive inhibition test showed that four of the 11 phages displayed foreign polypeptides had no common characteristics,which might be non-specific binding.All of the seven cloned amino acid sequences contained the same structure(-WP-Q-)of proline(P),tryptophan(W)and glutamic acid(Q),with an inhibition rate of more than 40%.Western blot analysis showed that the selected phage could be specifically recognized by anti Rh(D)serum,and its antigenicity was similar to that of Rh(D)protein.【Conclusion】Using anti-Rh(D)monoclonal antibody to screen the phage random peptide library,the Rh(D)antigen mimotope with(-WP-Q-)structure can be obtained.

关 键 词：RHD抗原 筛选效率 序列分析 医学鉴定 

分 类 号：R457.1[医药卫生—治疗学]