运用噬菌体随机肽库分析系统性红斑狼疮的Sm抗原模拟表位  

The study of Sm antigen mimic epitope of SLE by 12 mer phage random peptide library

作　　者：谢红付[1] 树叶[1] 张慧[1] 施为[1] 冯浩[1] 

机构地区：[1]中南大学附属湘雅医院皮肤科,湖南长沙410000

出　　处：《临床皮肤科杂志》2004年第10期600-603,共4页Journal of Clinical Dermatology

基　　金：湖南省自然科学基金项目(01JJY2015)

摘　　要：目的:应用噬菌体呈现技术分析系统性红斑狼疮(SLE)Sm抗原的模拟表位。方法:以鼠单克隆抗Sm抗体为筛选配基,对噬菌体随机12肽库进行3轮亲和筛选。分离10个噬菌体克隆进行Dot-ELISA和双抗体夹心ELISA鉴定。将阳性克隆进行DNA序列测定,并将混合阳性克隆与患者血清反应。结果:经3轮筛选,每轮筛选的投入与产出比逐轮升高,显示良好的富集效果。两种ELISA鉴定9个噬菌体克隆具有特异性。经过DNA测序得到6个不同的序列,推导出保守的氨基酸序列为RR/PR/IS。混合克隆诊断sLE血清的敏感性是51.72％,特异性是100.00％,结论:噬菌体呈现技术能够分析Sm抗原的模拟表位,将有利于研制表位水平的诊断试剂。Objective: To analyze Sm antigen mimic epitopes of SLE by phage display technique. Methods: Anti-SM monoclonal antibody was used to immunoscreen 12 mer phage random peptide library and 10 clones were selected after 3 rounds of biopanning. The reactivity of the 10 clones bound to anti-SM monoclonal antibody was examined by Dot-ELISA and DAS-ELISA. Positive clones were sequenced. The assembled positive clones were reacted with patient serum. Results: By screening, the ratio of output/input increased markedly, which showed an efficient enrichment effect. The specificity of 9 clones was verified by Dot-ELISA and DAS ELISA. And 6 different sequences were present, which deduced possible conservative aminoacid series-namely RR/PR/IS. The sensitivity of serum experiment was 51.72% and the specificity was 100.00%. Conclusion: Phage display technique could analyze SM antigen mimic epitope, which can be helpful to develop diagnostic reagent on the epitope level.

关 键 词：红斑狼疮 系统性 噬菌体呈现技术 抗原 SM 表位 

分 类 号：R593.241[医药卫生—内科学] R392.11[医药卫生—临床医学]