Sensitive Chemiluminescence Immunoassay by Capillary Electrophoresis with Gold Nanoparticles
Sensitive Chemiluminescence Immunoassay by Capillary Electrophoresis with Gold Nanoparticles
复制标题
金纳米颗粒毛细管电泳灵敏化学发光免疫分析
DOI：
10.1021/ac103166n
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发表时间：
2011-02-01
影响因子：
7.4
通讯作者：
Rei, Shu-wei
中科院分区：
文献类型：
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作者：
Liu, Yan-ming;Mei, Lin;Rei, Shu-wei
本技术说明描述了一种与毛细管电泳（基于 CE 的 CL-IA）联用的新型化学发光免疫测定法，采用金纳米粒子 (AuNP) 技术进行生物分子测定。 AuNPs因其对鲁米诺和过氧化氢的CL反应具有优异的催化作用而被用作蛋白质标记试剂。 AuNPs 与抗体 (Ab) 缀合形成标记抗体 (Ab*)，然后 Ab* 与抗原 (Ag) 连接，通过非竞争性免疫反应产生 Ab*-Ag 复合物。在优化条件下，将过量 Ab* 和 Ab*-Ag 复合物的混合物进行基线分离并在 5 分钟内检测到 1。这一新方案以人免疫球蛋白 G (IgG) 作为目标分子进行了评估。 IgG的校准曲线在0.008-5μg/mL范围内，相关系数为0.995。 IgG的检测限(S/N=3)为1.14×10(-3)μg/mL(7.1pmol/L,0.39amol)。所提出的 AuNPs 增强的基于 CE 的 CL-IA 方法已成功应用于患者人血清中 IgG 的定量。证明该方法可发展成为一种新型、灵敏的生化分析技术。
This technical note describes a new chemiluminescence immunoassay hyphenated to capillary electrophoresis (CE-based CL-IA) with gold nanoparticles (AuNPs) technique for biological molecules determination. AuNPs were used as a protein label reagent in the light of its excellent catalytic effect to the CL reaction of luminol and hydrogen peroxide. AuNPs conjugate with antibody (Ab) to form tagged antibody (Ab*), and then Ab* link to antigen (Ag) to produce an Ab*-Ag complex by a noncompetitive immunoreaction. The mixture of the excess Ab* and the Ab*-Ag complex was baseline separated and detected 1 within 5 min under the optimized conditions. This new protocol was evaluated with human immunoglobulin G (IgG) as the target molecule. The calibration curve of IgG was in the range of 0.008-5 mu g/mL with a correlation coefficient of 0.995. The detection limit (S/N = 3) of IgG was 1.14 x 10(-3) mu g/mL (7.1 pmol/L, 0.39 amol). The proposed AuNPs enhanced CE-based CL-IA method was successfully applied for the quantification of IgG in human sera from patients. It proves that the present method could be developed into a new and sensitive biochemical analysis technique.