Discovery of a polystyrene binding peptide isolated from phage display library and its application in peptide immobilization.
Discovery of a polystyrene binding peptide isolated from phage display library and its application in peptide immobilization.
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DOI：
10.1038/s41598-017-02891-x
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发表时间：
2017-06-01
影响因子：
4.6
通讯作者：
Qiu Z
中科院分区：
文献类型：
--
作者：
Qiang X;Sun K;Xing L;Xu Y;Wang H;Zhou Z;Zhang J;Zhang F;Caliskan B;Wang M;Qiu Z
噬菌体肽段展示技术是一种发现各种靶向特异性配体的有效技术。然而，靶不相关的肽往往可以获得，并导致模糊的结果。在我们实验室中，针对不同的靶标反复分离肽PB-TUP，并且我们对PB-TUP噬菌体进行了研究，以研究它们的结合特性和繁殖速率。ELISA和噬菌体回收试验表明，PB-TUP噬菌体与聚苯乙烯固体表面的亲和力明显上级对照噬菌体克隆。在本研究中，排除了一些偶然结合，如封闭剂和二抗的非特异性结合。对所选噬菌体克隆的增殖率测定表明，PB-TUP噬菌体的生长率并不上级对照噬菌体。PB-TUP与聚苯乙烯的结合具有浓度依赖性，并随溶液pH值的变化而变化。分子模拟结果表明，PB-TUP与聚苯乙烯的结合可能与其稳定的α-螺旋和β-转角结构有关。将PB-TUP序列融合到肽P2的N-末端，并且融合肽显著增加了与聚苯乙烯的结合亲和力。该融合肽还能增强肽P2与人脐静脉内皮细胞（HUVEC）的粘附能力。聚苯乙烯结合肽的加入为肽的固定化提供了一种方便的方法。
Phage peptide display is a powerful technique for discovery of various target-specific ligands. However, target-unrelated peptides can often be obtained and cause ambiguous results. Peptide PB-TUP has been isolated repeatedly in our laboratory on different targets and we conducted a research on PB-TUP phage to investigate their binding properties and rate of propagation. ELISA and phage recovery assay demonstrated that PB-TUP phage had a significant superior affinity to polystyrene solid surface compared with control phage clones. In this study, some incidental bindings are excluded like blocking agents and non-specific binding of secondary antibodies. Propagation rate assays of the selected phage clones showed that the growth rate of PB-TUP phage was not superior to the control phages. Furthermore, the binding of PB-TUB to polystyrene was concentration dependent and varied with solution pH. Molecular modeling revealed that stable structures of α-helix and β-turn may contribute to the binding of PB-TUP to polystyrene plate. The PB-TUP sequence was fused to the N-terminus of peptide P2 and the fusion peptide significantly increased the binding affinity to polystyrene. The fusion peptide also enhanced the cell adhesion ability of peptide P2 with human umbilical vein endothelial cell (HUVEC). The addition of the polystyrene binding peptide provided a convenient method for peptide immobilization.