Fluorescein isothiocyanate linked immunoabsorbent assay based on surface-enhanced resonance Raman scattering
Fluorescein isothiocyanate linked immunoabsorbent assay based on surface-enhanced resonance Raman scattering
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基于表面增强共振拉曼散射的异硫氰酸荧光素联免疫吸附测定
DOI：
10.1021/ac702497t
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发表时间：
2008-04-15
影响因子：
7.4
通讯作者：
Ozaki, Yukihiro
中科院分区：
文献类型：
--
作者：
Han, Xiao X.;Cai, Lin J.;Ozaki, Yukihiro
以异硫氰酸荧光素（FITC）为拉曼探针，建立了一种简单、灵敏的表面增强共振拉曼散射免疫分析方法。首次报道了基于SERRS的微量滴定板底部的免疫测定。本研究采用了酶联免疫吸附法（ELISA）作为主要的前处理方法。在该方法中，在抗原包被、封闭、抗体添加和胶体银染色的几个连续步骤之后测量FITC的SERRS光谱。人免疫球蛋白G（IgG）和FITC-抗人IgG用于免疫反应。所提出的方法对于免疫测定具有几个优点。首先，我们可以通过与抗体连接的FITC分子的SERRS信号的强度来确定抗原的浓度，而无需酶反应，因此该过程简单且节省试剂。第二，可以直接从微量滴定板底部的银聚集体获得FITC的SERRS光谱，而无需置换。第三，通过使用FITC的SERRS，本方法在0.2ng/mL的浓度下足以检测抗原，这与ELISA相当。结果表明，建议的SERRS为基础的免疫分析可能有很大的潜力，作为一个高灵敏度和高通量的免疫分析。
By using fluorescein isothiocyanate (FITC) as a Raman probe, we have developed a simple and sensitive method for an immunoassay based on surface-enhanced resonance Raman scattering (SERRS). For the first time, a SERRS-based immunoassay on the bottom of a microtiter plate is reported. We have applied the main pretreatment method of enzyme-linked immunoabsorbent assay (ELISA) to the present study. In this method, SERRS spectra of FITC are measured after several continuous steps of antigen coating, blocking, antibody adding, and colloidal silver staining. Human immunoglobulin G (IgG) and FITC-antihuman IgG are used for the immunoreaction. The proposed method has several advantages for immunoassay. First, we can determine the concentration of antigens via the intensity of a SERRS signal of FITC molecules that are attached to antibodies without an enzyme reaction, and thus the process is simple and reagent saving. Second, one can obtain SERRS spectra of FITC directly from silver aggregates on the bottom of a microtiter plate without displacement. Third, by using SERRS of FITC, the present method is sensitive enough to detect antigens at the concentration of 0.2 ng/mL, which is comparable to ELISA. Results are presented to demonstrate that the proposed SERRS-based immunoassay may have great potential as a high-sensitivity and high-throughout immunoassay.