利用噬菌体肽库技术筛选卵巢特异的VEGFR3亲和肽

目的从噬菌体12肽库中筛选VEGFR3胞外区蛋白高亲和肽作为卵巢癌淋巴管上皮细胞潜在靶向载体。方法用噬菌体展示固相结合法,生物淘选与扩增。经过4个循环的筛选,利用ELISA法鉴定噬菌体单克隆的亲和性,测定亲和力高的阳性噬菌体融合蛋白的DNA序列,竞争法ELISA检测优势克隆与靶蛋白的特异性。结果经过4轮筛选,噬菌体出现良好的富集,选择8个阳性克隆测序,经测序5个克隆插入短肽是(WHGSLKQNLWWY),竞争性ELISA显示其与VEGFR3具有良好的亲合性,并能被VEGF-D分子抑制。结论噬菌体12肽库筛选的VEGFR3高亲和多肽(WHGSLKQNLWWY),可望成为靶向载体构建的结构基础。

Screening the peptide high-binding VEGF receptor 3 with phage-displayed random peptide library

Objective To get a high affinity peptide of vascular endothelial growth factor receptor 3 (VEGFR3) as a potent carrier targeted to lymphangiogenesis of ovarian cancer via the technology of phage display. Methods Solid-phase was panned with direct VEGFR3 extracellular protein coating, then the unbound phage was washed away and the eluted phage was amplified. The positive phage clones were identified by ELISA and sequenced, and the affinity and specialty were identified by competent ELISA. Results After four-round bio-panning, the enriched positive phage clones were identified by ELISA. Eight positive phage clones were sequenced and 5 were consensus (WHGSLKQNLWWY). The short peptide displayed on screened positive phage could bind specifically to VEGFR3, and the binding could be inhibited by natural antibody VEGF-D. Conclusion The phage clone (phage-WHGSLKQNLWWY) obtained via bio-panning of peptide library has a high affinity with VEGF receptor 3. The peptide could be a potent carrier targeted to VEGFR3.