Department of Chemistry and Institute for Advanced Study, Nanchang University, Nanchang 330031, China.
Anal Chim Acta. 2012 Aug 6;737:22-8. doi: 10.1016/j.aca.2012.05.043. Epub 2012 Jun 9.
Small molecules or analytes present at low concentrations are difficult to detect directly using conventional surface plasmon resonance (SPR) techniques because only small changes in the refractive index of the medium are typically induced by the binding of these analytes. Here, we present an amplification technique using core-shell Fe(3)O(4)@Au magnetic nanoparticles (MNPs) for an SPR bioassay. To evaluate this amplification effect, a novel SPR sensor based on a sandwich immunoassay was developed to detect α-fetoprotein (AFP) by immobilizing a primary AFP antibody (Ab(1)) on the surface of a 3-mercapto-1-propanesulfonate/chitosan-ferrocene/Au NP (MPS/CS-Fc/Au NP) film employing Fe(3)O(4)@Au-AFP secondary antibody conjugates (Fe(3)O(4)@Au-Ab(2)) as the amplification reagent. The stepwise fabrication of the biosensor was characterized using UV-vis spectroscopy, electrochemical impedance spectroscopy, and cyclic voltammetry. A calibration curve of Fe(3)O(4)@Au-Ab(2) conjugates amplification for AFP detection was obtained to yield a correlation in the range of 1.0-200.0 ng mL(-1) with a detection limit of 0.65 ng mL(-1), and a significant increase in sensitivity was therefore afforded through the use of Fe(3)O(4)@Au-Ab(2) conjugates as an amplifier. This magnetic separation and amplification strategy has great potential for the detection of other biomolecules of interest with low interference and high sensitivity by changing the antibody label used in the Fe(3)O(4)@Au-antibody conjugates.
小分子或浓度较低的分析物很难直接用传统的表面等离子体共振 (SPR) 技术检测,因为这些分析物的结合通常只会引起介质折射率的微小变化。在这里,我们提出了一种使用核壳 Fe(3)O(4)@Au 磁性纳米粒子 (MNP) 的放大技术用于 SPR 生物分析。为了评估这种放大效应,我们开发了一种基于三明治免疫测定的新型 SPR 传感器,通过将α-胎蛋白 (AFP) 的一级抗体 (Ab(1)) 固定在 3-巯基-1-丙磺酸/壳聚糖-二茂铁/Au NP (MPS/CS-Fc/Au NP) 膜的表面上,利用 Fe(3)O(4)@Au-AFP 二级抗体缀合物 (Fe(3)O(4)@Au-Ab(2)) 作为放大试剂,来检测 AFP。采用紫外可见光谱、电化学阻抗谱和循环伏安法对生物传感器的逐步制备进行了表征。获得了 Fe(3)O(4)@Au-Ab(2) 缀合物放大 AFP 检测的校准曲线,在 1.0-200.0 ng mL(-1) 范围内具有相关性,检测限为 0.65 ng mL(-1),因此通过使用 Fe(3)O(4)@Au-Ab(2) 缀合物作为放大器,灵敏度得到了显著提高。这种磁分离和放大策略具有很大的潜力,通过改变 Fe(3)O(4)@Au-抗体缀合物中使用的抗体标记,可以实现对其他感兴趣的生物分子的低干扰、高灵敏度检测。