利用噬菌体随机15肽库确定SLE相关多肽的研究  

The study of SLE associated peptide by phage-display random 15 amino acid peptide library

作　　者：施明[1] 柯屾 陶永涛[1] 唐福林[3] 张奉春[3] 姚志建 沈倍奋[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850 [2]第二军医大学 [3]北京协和医院 [4]北京北方基因中心

出　　处：《中华微生物学和免疫学杂志》2001年第3期275-277,共3页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金重点项目! (3 983 0 3 3 0 ) ;国家"863"计划资助项目! (10 2 0 9 0 3 0 4)

摘　　要：目的　利用噬菌体随机 15肽库确定SLE相关多肽。方法　从系统性红斑狼疮 (SLE)病人血清中纯化抗dsDNA多克隆抗体 ,以此为筛选配基 ,对噬菌体随机 15肽库进行亲和筛选。结果经 3轮筛选 ,每轮的投入 /产出比逐轮升高 ,提示筛选具有良好的富集效果。第 3轮筛选后选其中 18个克隆进行结合试验 ,结果显示有 6个仅与SLE病人血清反应而不与正常人血清反应 ,经DNA序列测定 ,发现其中 5个克隆序列相同 ,通过该序列推出插入的外源短肽序列 ,合成并纯化该肽段。用此肽段分别与 2 0例SLE病人及 2 0例正常人血清反应 ,结果差异有显著性。结论　该肽段可能是与SLE相关的抗原多肽。Objective The study of SLE related peptide by phage-display random 15 amino acid peptide library. Methods Anti-dsDNA antibody was purified from the serum of the SLE patients. The antibody was used as the ligand for biopanning of a phage-displayed random peptide library of 15 amino acid residues. Results After three rounds of screening, the ratio of output to input was increased. It indicated the efficient enrichment. At the third round of screening, eighteen clones were selected for binding test with the sera of patients and of normal people. The results showed that six clones could react with the patients′ serum but not the serum of normal people. By DNA sequencing, it was confirmed that in five clones the fragments inserted had the identical sequence. The peptide was synthesized from the deduced sequence. There is significant difference between the reactions of patients′ and normal people′s serum with the synthetic peptide. Conclusion The peptide was possibly associated with SLE antigens.

关 键 词：噬菌体 随机肽库 抗体 系统性红斑狼疮 发病机制 

分 类 号：R593.241[医药卫生—内科学]