噬菌体随机肽库中B型钠尿肽结合肽的筛选及鉴定

Screening and identification of polypeptides specifically binding to human B type natriuretic peptide from 12TM phage display peptide library

目的利用噬菌体表面展示技术筛选与人B型钠尿肽(BNP)结合的特异性单链噬菌体多肽,并鉴定多肽与BNP的结合力。方法以BNP为靶分子,筛选噬菌体线性12肽库,ELISA鉴定阳性克隆。结果经4轮淘选后,随机挑选16个噬菌体克隆,其中10个克隆与BNP有较强的结合能力,序列测定得到5种氨基酸序列;阳性克隆与BNP结合的剂量依赖实验结果提示淘选获得的多肽能与BNP特异性结合。结论经筛选得到的12肽能与BNP特异性结合,为下一步多肽的表达纯化及制备BNP检测试剂盒提供了重要的基础。

Objective To identify and characterize the polypeptides specifically binding to human B type natriuretic peptide （BNP） screened from 12TM phage display peptide library. Methods The BNP-binding peptides were screened from 12TM phage display peptide library and identified by ELISA. Results After 4 rounds of screening, 10 of the 16 phage clones were identified as the positive clones which could bind to BNP. Five amino acid sequences were obtained in the 10 positive clones. Dose-dependent ELISA results demonstrated that the screened polypeptides could specifically bind to BNP. Conclusion These screened polypeptides can bind specifically to BNP, which provides a basis for further research on expression and purification of anti-BNP polypeptides and the development of the detection kit of BNP.