Peelen Dora, Kodoyianni Voula, Lee Jieun, Zheng Ting, Shortreed Michael R, Smith Lloyd M
Department of Chemistry, 1101 University Avenue, University of Wisconsin, Madison, Wisconsin 53706-1396, USA.
J Proteome Res. 2006 Jul;5(7):1580-5. doi: 10.1021/pr050467e.
Aldehyde-terminated self-assembled monolayers (SAMs) on gold surfaces were modified with proteins and employed to capture intact living cells through specific ligand-cell surface receptor interactions. In our model system, the basic fibroblast growth factor (bFGF) binding receptor was targeted on baby hamster kidney (BHK-21) cells. Negative control and target proteins were immobilized on a gold surface by coupling protein primary amines to surface aldehyde groups. Cell-binding was monitored by phase contrast microscopy or surface plasmon resonance (SPR) imaging. The specificity of the receptor-ligand interaction was confirmed by the lack of cell binding to the negative control proteins, cytochrome c and insulin, and by the disruption of cell binding by treatment with heparitinase to destroy heparan sulfate which plays an essential role in the binding of bFGF to FGF receptors. This approach can simultaneously probe a large number of receptor-ligand interactions in cell populations and has potential for targeting and isolating cells from mixtures according to the receptors expressed on their surface.
在金表面的醛基终止自组装单分子层（SAMs）用蛋白质进行修饰，并通过特定的配体 - 细胞表面受体相互作用用于捕获完整的活细胞。在我们的模型系统中，碱性成纤维细胞生长因子（bFGF）结合受体以幼仓鼠肾（BHK - 21）细胞为靶点。通过将蛋白质伯胺与表面醛基偶联，将阴性对照蛋白和靶蛋白固定在金表面。通过相差显微镜或表面等离子体共振（SPR）成像监测细胞结合情况。阴性对照蛋白细胞色素c和胰岛素未出现细胞结合现象，以及用肝素酶处理破坏硫酸乙酰肝素（其在bFGF与FGF受体结合中起重要作用）后细胞结合被破坏，证实了受体 - 配体相互作用的特异性。这种方法可以同时探测细胞群体中大量的受体 - 配体相互作用，并且具有根据细胞表面表达的受体从混合物中靶向和分离细胞的潜力。
