噬菌体随机肽库技术筛选抗CCR5单克隆抗体的识别表位  

Screening of Antigenic Determinants Recognized by Monoclonal Antibody against CC Chemokine Receptor 5 from A 12-mer Random Peptide Library

作　　者：郭海萍[1] 周至威[1] 叶惠荣[1] 黄秋月[1] 

机构地区：[1]广州医学院病原生物学教研室,广州510182

出　　处：《中国生物制品学杂志》2010年第10期1071-1073,共3页Chinese Journal of Biologicals

基　　金：广州市教育局市属高校科技社科项目(61046)

摘　　要：目的利用噬菌体随机肽库技术筛选趋化因子受体5(CCchemokine receptor 5,CCR5)单克隆抗体的识别表位。方法用抗CCR5单克隆抗体筛选噬菌体随机12肽库,采用双抗体夹心ELISA法鉴定噬菌体阳性克隆,挑取阳性克隆测定DNA序列,推导其氨基酸序列并进行同源性分析。结果从噬菌体随机12肽库中筛选出30株可与抗CCR5单抗特异结合的噬菌体克隆,其中多数克隆呈现核心序列HY-TC-SS-XX-P/FYS-X,该序列与CCR5的一级序列ECL2具有一定的同源性。结论 HY-TC-SS-XX-P/FYS-X序列是抗CCR5单抗的识别表位。Objective To screen the antige nic determinants recognized by monoclonal antibody（McAb） against CC chemokine receptor 5（CCR5）from a 12-mer random peptide library. Methods A 12-mer random peptide library was screened by using McAb against CCR5, and the positive clones were indentified by double antibody sandwich ELISA and subjected to DNA sequencing, based on which their amino acid sequences were deduced and analyzed for homology. The binding character of the obtained short phage peptide to the McAb was identified by ELISA. Results Thirty phage clones specifically binding to McAb against CCR5 were screened from the 12-mer random peptide library, in most of which core sequence HY-TC-SS-XX-P / FYS-X were found, with a certain homology to the primary sequence ECL2 of CCR5. Conclusion The sequence HY-TC-SS-XX-P / FYS-X was the antigenic determinants recognized by McAb against CCR5.

关 键 词：噬菌体随机12肽库 受体 CCR5 抗体 单克隆 识别表位 

分 类 号：Q78[生物学—分子生物学] R392.33[医药卫生—免疫学]