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目的获取与CC趋化因子受体9(CCR9)具有亲和力的特定短肽。方法以CCR9的第2个胞外环为靶蛋白,反复筛选Ph.D.-12噬菌体展示肽库获得阳性克隆,通过ELISA测定噬菌体阳性克隆短肽与CCR9的亲和力,共聚焦显微镜检测短肽与CCR9高表达细胞MOLT4结合能力。结果经过3轮淘选、富集,确定了8个噬菌体克隆有插入序列,其中克隆C-4与靶蛋白具有较高亲和力,表达的短肽P1(VHWDFRQWWQPS)可抑制相应C-4克隆与靶蛋白的结合,并可与CCR9高表达的MOLT4细胞结合。结论通过噬菌体肽库筛选出了能与细胞表面分子CCR9结合的短肽序列VHWDFRQWWQPS。
Objective To obtain specific short peptides with affinity for CC chemokine receptor 9 (CCR9). Methods The second extracellular loop of CCR9 was used as the target protein and the Ph.D.-12 phage display peptide library was screened repeatedly to obtain the positive clones. The affinity of the phage positive clones with CCR9 was determined by ELISA. The confocal microscopy CCR9 Highly Expressed Cell MOLT4 Binding Capacity. Results After 3 rounds of panning and enrichment, 8 phage clones were identified as insertions. Clone C-4 showed high affinity with the target protein. The expressed short peptide P1 (VHWDFRQWWQPS) inhibited the interaction between the corresponding C-4 clone and the target Protein and bind to MOLT4 cells with high CCR9 expression. Conclusion The peptide sequence VHWDFRQWWQPS, which can bind to the cell surface molecule CCR9, was screened by the phage peptide library.