中文摘要：

目的：利用噬菌体展示技术，筛选VEGF受体3（VEGFR3）胞外区蛋白高亲和噬菌体融合肽，并测量其亲和常数。方法：用噬菌体展示固相结合法，经过4个循环的筛选，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，并通过非竞争性ELISA固项法，得到融合肽与VEGFR3的抗原抗体结合反应曲线，计算融合肽的亲和常数。结果：4轮亲和筛选，噬菌体出现良好的富集，8个阳性克隆中,经测序有4个克隆序列相同（phage—WHGSLKQNLWWY）；竞争性ELISA显示噬菌体融合肽与VEGFR3具有良好的亲合性，并能被VEGF—D分子抑制，融合肽的亲和常数为（1．505±0．184）×10^7M^-1。结论：噬菌体12肽库筛选的VEGFR3高亲和融合肽（phage—WHGSLKQNLWWY）可为多肽分子的改造及靶向药物的开发提供依据．

英文摘要：

Objective： To get a high affinity fused - peptide of vascular endothelial growth factor receptor 3 via the technology of phage display and measure the functional affinity between humanized VEGF receptor 3（VEGF R3） and the fused peptide. Methods： 1. Direct VEGF R3 exta - cellular protein coating, solid - phase panning. 2. Washed away the unbound phage and amplified the eluted phage. 3. Identified the positive phage clones by ELISA and sequenced them. 4. Identified the affinity and sepeciality by competent ELISA. 5. Assess the functional affinity of engineered peptide and VEGF - D with non - competitive ELISA. Results. After four rounds biopanning, the enriched positive phage clones were identified by ELISA. 8 positive phage clones were sequenced and 4 were consensus （WHGSLKQNLWWY）, the short peptide displayed on screened positive phage could bind specifically to VEGF R3, and the binding could be inhibited by natural antibody VEGF - D. the affinity of fused peptide was （2. 47±1.46） × 107/M. Conclusion. The phage clone （phage - WHGSLKQNLWWY） got via biopanning of peptide library has a high affinity with VEGF R3. The fused peptide can be a potent cartier targeted to VEGF R3.