sFcγRⅡb蛋白结合肽的筛选与鉴定  

Screening and identification of sFcγRⅡb binding peptide

作　　者：徐臻臻 刘利鹏[1,2] 张艳芬 崔哲 毕克维[1,2] 刘中成 XU Zhenzhen;LIU Lipeng;ZHANG Yanfen;CUI Zhe;BI Kewei;LIU Zhongcheng(College of Pharmaceutical Sciences, Hebei University, Baoding 071002, China;Key Laboratory of Pharmaceutical Quality Control of Hebei Province, Baoding 071002, China;Office of Science and Technology, Hebei University, Baoding 071002, China)

机构地区：[1]河北大学药学院,河北保定071002 [2]河北省药物质量分析与控制重点实验室,河北保定071002 [3]河北大学科学技术处,河北保定071002

出　　处：《河北大学学报（自然科学版）》2019年第4期398-403,共6页Journal of Hebei University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(81202338);河北省自然科学基金资助项目(H2016201121);河北省高等学校科学技术研究项目(ZD2017010)

摘　　要：FcγRⅡb是免疫球蛋白G受体(FcγR)中唯一的抑制型受体,在免疫反应的负性调节方面发挥重要作用.为了筛选sFcγRⅡb的蛋白结合肽,以重组sFcγRⅡb蛋白为靶分子,采用噬菌体肽库展示技术对sFcγRⅡb结合肽进行筛选.利用ELISA鉴定每轮洗脱噬菌体与sFcγRⅡb蛋白亲和力,经过4轮筛选,挑取40个噬菌体克隆进行序列测定,获得28种不同的12肽序列.经ELISA法鉴定噬菌体与sFcγRⅡb蛋白结合活性,得到sFcγRⅡb蛋白高特异性、高亲和力结合肽FHKMPWYMSMYY,为进一步研究FcγRⅡb的作用机制和探索结合肽的功能提供实验基础.FcγR Ⅱ b is the only inhibitory receptor of the IgG Fc receptor (FcγR) and plays an important role in the negative regulation and immune responses.In order to screen the binding peptide of sFcγR Ⅱ b , the recombinant sFcγR Ⅱ b protein was used as the target molecule, and the sFcγR Ⅱ b binding peptide was screened by phage peptide library display technology.The affinity of each eluted phage to sFcγR Ⅱ b protein was identified by ELISA.40 phage clones were screened after four rounds screening, and 28 different 12-residue peptides were obtained through translation of DNA sequencing data.The affinities between phages and sFcγR Ⅱ b protein were estimated and confirmed by ELISA analysis, and the results showed that the deduced peptide sequence, FHKMPWYMSMYY was the highest specificity and affinity to sFcγR Ⅱ b protein. Furthermore, this study may lay a foundation for the further research on the mechanism of FcγR Ⅱ b and the function of binding peptide.

关 键 词：sFcγRⅡb 噬菌体肽库展示技术 结合肽 

分 类 号：Q816[生物学—生物工程]