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.
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.
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