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一种新颖的低成本且易于开发的功能化平台。案例研究:基于适配体的表面等离子体共振法检测凝血酶。

A novel low-cost and easy to develop functionalization platform. Case study: aptamer-based detection of thrombin by surface plasmon resonance.

机构信息

International Centre of Biodynamics, Intrarea Portocalelor 1B, 060101 Bucharest, Romania.

出版信息

Talanta. 2010 Mar 15;80(5):2157-64. doi: 10.1016/j.talanta.2009.11.023. Epub 2009 Nov 12.

Abstract

A novel low-cost platform to assess biomolecular interactions was investigated using surface plasmon resonance and an aptamer-based assay for thrombin detection. Gold SPR surface functionalized with a carboxylated cross-linked BSA film (cBSA) and commercially available carboxymethylated dextran chip (CM5) were used as immobilization platforms for the thrombin binding aptamer. The high end commercial instrument Biacore 3000 and a custom made FIA set-up involving TI Spreeta sensor (TSPR2K23) were used to assess different concentrations of thrombin within the range 0.1-150 nM both in buffer and in a complex matrix (plasma) using the obtained aptasensors. Based on data derived from both CM5 and cBSA platforms, the cBSA aptasensor exhibited good selectivity, stability and regeneration ability, both in buffer and in complex matrices (plasma), comparable with CM5.

摘要

研究了一种新型的低成本平台,该平台使用表面等离子体共振和基于适体的测定法来评估生物分子相互作用,用于凝血酶检测。金 SPR 表面用羧基化交联 BSA 膜 (cBSA) 和市售的羧甲基化葡聚糖芯片 (CM5) 进行功能化,作为凝血酶结合适体的固定化平台。高精密商业仪器 Biacore 3000 和一个定制的 FIA 装置,涉及 TI Spreeta 传感器 (TSPR2K23) ,用于评估不同浓度的凝血酶在 0.1-150 nM 范围内的缓冲液和复杂基质(血浆)中,使用获得的适体传感器。基于来自 CM5 和 cBSA 平台的数据,cBSA 适体传感器在缓冲液和复杂基质(血浆)中均表现出良好的选择性、稳定性和再生能力,与 CM5 相当。

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