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Specific and reversible immobilization of histidine-tagged proteins on functionalized silicon nanowires.

机译:组氨酸标签蛋白在功能化硅纳米线上的特异性和可逆固定。

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摘要

Silicon nanowire (Si NW)-based field effect transistors (FETs) have shown great potential as biosensors (bioFETs) for ultra-sensitive and label-free detection of biomolecular interactions. Their sensitivity depends not only on the device properties, but also on the function of the biological recognition motif attached to the Si NWs. In this study, we show that SiNWs can be chemically functionalized with Ni:NTA motifs, suitable for the specific immobilization of proteins via a short polyhistidine tag (His-tag) at close proximity to the SiNW surface. We demonstrate that the proteins preserve their function upon immobilization onto SiNWs. Importantly, the protein immobilization on the Si NWs is shown to be reversible after addition of EDTA or imidazole, thus allowing the regeneration of the bioFET when needed, such as in the case of proteins having a limited lifetime. We anticipate that our methodology may find a generic use for the development of bioFETs exploiting functional protein assays because of its high compatibility to various types of NWs and proteins.
机译:基于硅纳米线(Si NW)的场效应晶体管(FET)作为生物传感器(bioFET)表现出巨大的潜力,可用于生物分子相互作用的超灵敏且无标记的检测。它们的敏感性不仅取决于器件的性能,而且取决于附着在Si NW上的生物识别基序的功能。在这项研究中,我们表明,SiNWs可以通过Ni:NTA基序进行化学功能化,适用于通过紧密靠近SiNW表面的短聚组氨酸标签(His-tag)对蛋白质进行特异性固定。我们证明蛋白质固定在SiNWs上时保留其功能。重要的是,显示出在添加EDTA或咪唑之后,将蛋白质固定在Si NW上是可逆的,从而在需要时允许bioFET再生,例如在蛋白质寿命有限的情况下。我们期望我们的方法论可以发现开发功能性蛋白分析的bioFET的通用方法,因为它与各种类型的NW和蛋白具有高度的兼容性。

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