期刊论文

中文

中国免疫学杂志

1000-484X

2015

31

11

1465-1471

10.3969/j.issn.1000-484X.2015.11.006

湖北省自然科学基金重点项目（2014CFA079） 国家自然科学基金（No.30972693,21473065）资助.

本校为第一机构

目的: 鉴定RA相关自身抗原COMP的一株特异性单克隆抗体15A11的表位特征。方法: 选用随机十二肽噬菌体库对mAb 15A11进行三轮筛选,随机挑取40个单噬菌斑,提取DNA,测序; ELISA检测每个噬菌体克隆与mAb 15A11结合的特异性; 通过ClustalW2对特异性结合的噬菌体展示的十二肽和COMP进行氨基酸序列比对,PyMOL分析一致氨基酸所在肽链的二级结构及表位氨基酸之间的距离,初步确定mAb 15A11的表位; 变性和非变性Western blot、EDTA螯合Ca~(2+)后ELISA分析以及合成多肽的ELISA实验进一步确定表位序列。结果: 得到的40个测序噬菌体中,共有5个噬菌体克隆,ELISA确定克隆1和克隆2与mAb 15A11特异性结合,其他克隆...

Objective： To identify the epitope of mAb15A11 which is specific against RA associated autoantigen cartilage oligomeric matrix protein（ COMP）. Methods： A filamentous phage library displaying random linear dodecapeptides was used to mapping the epitope of mAb15A11. After three rounds of screenings,40 phage clones were selected at random and sequenced. The specificity of phages was confirmed by enzyme immunoassays. Homology search by Clustal W2 and structure analysis by Py Mol to identified the epitope amino acid sequence. Western blot analys...