sFcγRⅡb蛋白结合肽的筛选与鉴定

doi:10.3969/j.issn.1000-1565.2019.04.011

河北大学学报(自然科学版) ›› 2019, Vol. 39 ›› Issue (4): 398-403.DOI: 10.3969/j.issn.1000-1565.2019.04.011

sFcγRⅡb蛋白结合肽的筛选与鉴定

徐臻臻^1,2,刘利鹏^1,2,张艳芬^2,3,崔哲^1,2,毕克维^1,2,刘中成^1,2

收稿日期:2018-10-03 出版日期:2019-07-25 发布日期:2019-07-25
通讯作者: 刘中成(1979—),男,河北抚宁人,河北大学教授,博士,主要从事分子药理学研究.E-mail:liuzc@hbu.edu.cn
作者简介:徐臻臻(1993—),女,河北沧州人,河北大学在读硕士研究生. E-mail:1317446609@qq.com
基金资助:
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)

Screening and identification of sFcγRⅡb binding peptide

XU Zhenzhen^1,2, LIU Lipeng^1,2, ZHANG Yanfen^2,3, CUI Zhe^1,2, BI Kewei^1,2, LIU Zhongcheng^1,2

1.College of Pharmaceutical Sciences, Hebei University, Baoding 071002, China; 2.Key Laboratory of Pharmaceutical Quality Control of Hebei Province, Baoding 071002, China; 3.Office of Science and Technology, Hebei University, Baoding 071002, China

Received:2018-10-03 Online:2019-07-25 Published:2019-07-25

摘要/Abstract

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.

关键词: sFcγRⅡb, 噬菌体肽库展示技术, 结合肽

Abstract: FcγRⅡb 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 sFcγRⅡb, the recombinant sFcγRⅡb protein was used as the target molecule, and the sFcγRⅡb binding peptide was screened by phage peptide library display technology.The affinity of each eluted phage to sFcγRⅡb 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 sFcγRⅡb 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 sFcγRⅡb protein. Furthermore, this study may lay a foundation for the further research on the mechanism of FcγRⅡb and the function of binding peptide.

Key words: sFcγRⅡb, phage display technology, binding peptides

中图分类号:

Q816

徐臻臻,刘利鹏,张艳芬,崔哲,毕克维,刘中成. sFcγRⅡb蛋白结合肽的筛选与鉴定[J]. 河北大学学报(自然科学版), 2019, 39(4): 398-403.

XU Zhenzhen, LIU Lipeng, ZHANG Yanfen, CUI Zhe, BI Kewei, LIU Zhongcheng. Screening and identification of sFcγRⅡb binding peptide[J]. Journal of Hebei University (Natural Science Edition), 2019, 39(4): 398-403.

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sFcγRⅡb蛋白结合肽的筛选与鉴定

Screening and identification of sFcγRⅡb binding peptide

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