CNRS UMR-8182, Institut de Chimie Moléculaire et des Matériaux d'Orsay, Equipe de Chimie Bioorganique et Bioinorganique, Univ Paris-Sud, Orsay, France.
Anal Bioanal Chem. 2013 Mar;405(8):2505-14. doi: 10.1007/s00216-012-6665-4. Epub 2013 Jan 15.
We developed a biosensor based on the surface plasmon resonance (SPR) method for the study of the binding kinetics and detection of human cellular prions (PrP(C)) using DNA aptamers as bioreceptors. The biosensor was formed by immobilization of various biotinylated DNA aptamers on a surface of conducting polypyrrole modified by streptavidin. We demonstrated that PrP(C) interaction with DNA aptamers could be followed by measuring the variation of the resonance angle. This was studied using DNA aptamers of various configurations, including conventional single-stranded aptamers that contained a rigid double-stranded supporting part and aptamer dimers containing two binding sites. The kinetic constants determined by the SPR method suggest strong interaction of PrP(C) with various DNA aptamers depending on their configuration. SPR aptasensors have a high selectivity to PrP(C) and were regenerable by a brief wash in 0.1 M NaOH. The best limit of detection (4 nM) has been achieved with this biosensor based on DNA aptamers with one binding site but containing a double-stranded supporting part.
我们开发了一种基于表面等离子体共振（SPR）方法的生物传感器，用于使用 DNA 适体作为生物受体研究人类细胞朊病毒（PrP(C)) 的结合动力学和检测。该生物传感器通过将各种生物素化的 DNA 适体固定在经链霉亲和素修饰的导电聚吡咯表面上形成。我们证明，通过测量共振角的变化，可以跟踪 PrP(C)与 DNA 适体的相互作用。这是通过使用具有各种构型的 DNA 适体来研究的，包括包含刚性双链支撑部分的常规单链适体和包含两个结合位点的适体二聚体。通过 SPR 方法确定的动力学常数表明，PrP(C)与各种 DNA 适体之间存在强烈的相互作用，这取决于它们的构型。SPR 适体对 PrP(C)具有高选择性，并且可以通过在 0.1 M NaOH 中短暂冲洗进行再生。基于具有一个结合位点但包含双链支撑部分的 DNA 适体的这种生物传感器已实现最佳检测限（4 nM）。
