Zhao Wenxiu, Yuan Hang, Xu Xing, Ma Lan
Life Science Division, Graduate School at Shenzhen, Tsinghua University, Shenzhen, P. R. China.
J Biomol Screen. 2010 Jul;15(6):687-94. doi: 10.1177/1087057110370997. Epub 2010 May 27.
The search for new receptor ligands is important in the study of embryonic stem (ES) cell differentiation processes. In this study, a novel peptide (HGEVPRFHAVHL) with a specific ability to bind with neural stem cells derived from rhesus monkey ES cells was successfully screened out using a Ph.D-12 peptide phage display library. High affinity and specificity of the HGE phage were shown in an enzyme-linked immunosorbent assay. The binding ability of the phage could be matched with that of a chemically synthesized peptide with a sequence identical to that displayed by the phage, indicating that this binding capability manifests a peptide sequence. Combined with quantum dots, the HGE peptide can be used as a direct tool to show optical imaging of specific binding on a single cell membrane. Further results of Western blot showed that the HGE peptide interacted with 48/34-kDa proteins on the membrane of neural stem cells. This work is the first time that a phage display technique has been applied in ES cell differentiation studies. The findings extend the utilization of a targeting agent for neural stem cells and can also be used as a research tool in studying other cell lineages derived from ES cells.
寻找新的受体配体在胚胎干细胞(ES细胞)分化过程的研究中具有重要意义。在本研究中,利用Ph.D-12肽噬菌体展示文库成功筛选出一种与恒河猴ES细胞来源的神经干细胞具有特异性结合能力的新型肽(HGEVPRFHAVHL)。酶联免疫吸附测定显示HGE噬菌体具有高亲和力和特异性。噬菌体的结合能力与化学合成的、序列与噬菌体展示序列相同的肽的结合能力相当,表明这种结合能力由肽序列决定。与量子点结合后,HGE肽可作为直接工具用于显示单细胞膜上特异性结合的光学成像。蛋白质印迹的进一步结果表明,HGE肽与神经干细胞膜上的48/34-kDa蛋白相互作用。这项工作首次将噬菌体展示技术应用于ES细胞分化研究。这些发现扩展了神经干细胞靶向剂的应用范围,也可作为研究ES细胞衍生的其他细胞谱系的研究工具。