Immunoassay readout method using extrinsic Raman labels adsorbed on immunogold colloids
Immunoassay readout method using extrinsic Raman labels adsorbed on immunogold colloids
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DOI：
10.1021/ac990616a
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发表时间：
1999-11-01
影响因子：
7.4
通讯作者：
Porter, MD
中科院分区：
文献类型：
--
作者：
Ni, J;Lipert, RJ;Porter, MD
描述了一种基于表面增强拉曼散射（Sers）的免疫分析读出方法。该方法利用SERS衍生的报告分子，是coimmobilized与生物特异性物种的金胶体上的信号。这一概念在双分析物夹心测定中得到证实，其中两种不同的抗体共价结合到固体基质上，特异性地从水性样品中捕获两种不同的抗原。捕获的抗原进而选择性地结合至其相应的检测抗体。检测抗体与用不同的拉曼报告分子标记的金胶体缀合，所述报告分子用作每种类型抗体的外部标记。通过报告分子的特征Sers光谱确定特异性抗原的存在。近红外二极管激光器被用来有效地激发Sers信号，同时最大限度地减少荧光干扰。我们表明，通过使用不同的标签，几乎没有光谱重叠，可以同时检测到两种不同的抗原种类。简要讨论了这一概念作为多个分析物的读出策略的潜力。
An immunoassay readout method based on surface-enhanced Raman scattering (SERS) is described. The method exploits the SERS-derived signal from reporter molecules that are coimmobilized with biospecific species on gold colloids. This concept is demonstrated in a dual-analyte sandwich assay, in which two different antibodies covalently bound to a solid substrate specifically capture two different antigens from an aqueous sample. The captured antigens in turn bind selectively to their corresponding detection antibodies. The detection antibodies are conjugated with gold colloids that are labeled with different Raman reporter molecules, which serve as extrinsic labels for each type of antibody. The presence of a specific antigen is established by the characteristic SERS spectrum of the reporter molecule. A near-infrared diode laser was used to excite efficiently the SERS signal while minimizing fluorescence interference. We show that, by using different labels with little spectral overlap, two different antigenic species can be detected simultaneously. The potential of this concept to function as a readout strategy for multiple analytes is briefly discussed.