噬菌体随机肽库筛选动脉粥样硬化相关多肽  

Screening of atherosclerosis related polypeptide from phage display peptide library

作　　者：肖智[1] 赵序雯 赵明[1] 杨高红 

机构地区：[1]南方医科大学病理生理学教研室,广东省休克微循环重点实验室 [2]广东省功能蛋白质组学重点实验室,广东广州510515 [3]广州市天河区林和街社区卫生服务中心,广东广州510620

出　　处：《细胞与分子免疫学杂志》2013年第5期485-487,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81071549);广东省教育厅人才引进专项经费(粤财教2009年279号)

摘　　要：目的利用噬菌体展示技术筛选动脉粥样硬化疾病相关多肽,并验证所筛选的阳性噬菌体以及合成蛋白片段结合活性。方法以动脉粥样硬化患者血清为靶分子,噬菌体12肽库亲和筛选与之结合的阳性噬菌体。经过3轮筛选,从滴度测定平板上随机挑取10个阳性克隆,ELISA鉴定其亲和性。提取阳性噬菌体单链DNA并测序得到12肽序列,通过生物信息学查找相似天然蛋白,合成24肽,最后鉴定其结合活性。结果 ELISA显示阳性噬菌体均与患者血清有较强的结合力,序列为GPRPPSAPNMPL,合成24肽也呈现出较高的结合活性。结论噬菌体展示可以获得动脉粥样硬化相关多肽序列,为该病的免疫研究奠定了基础。Objective To screen atherosclerosis （AS） related polypeptide from phage display 12-peptide library, and verify the binding activity of selected positive phages and synthetic protein fragment. Methods We collected plasma from AS patients as target for biopanning against phage-displayed 12-peptide library. After 3 rounds of screening, l0 positive phages were picked up and the binding activity was proved by ELISA. Single-stranded DNA of the phages were purified and sequenced, and a similar polypeptide was synthesized to test the bJnding activity to AS patients plasma. Results Selected phages significantly bound to AS patients＇ plasma. Five of ten phages had GPRPPSAPNMPL sequence. Corresponding synthetic polypeptide also showed a high binding activity to AS patient plasma. Conclusion AS-related polypeptide can be obtained by phage display, which facilitates the research into the immune mechanism of AS.

关 键 词：噬菌体肽库 动脉粥样硬化 多肽合成 

分 类 号：Q74[生物学—分子生物学] R392-33[医药卫生—免疫学]