新型非催化质子免疫吸附测定放大机制

Aditya Saran , Shuai Hou , Yinglu Ji
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摘要

信号的放大取决于酶/催化剂。酶的活性、特异性、pH 值条件、酶连接物、稳定性、储存和效率等都是影响检测结果的重要因素。我们的目标是在没有酶/催化剂辅助的情况下放大信号,从而使该策略可用于包括免疫测定在内的各种信号检测。由于银纳米晶体(Ag NCs)在 ∼ 400 nm 处具有很强的等离子体波段,因此我们采用自核生长的银纳米晶体(Ag NCs)作为发色剂。在 6.10 mM CTAB 和 0.61 mM AgNO3 存在下的 140 秒反应时间内,0.57 mM 抗坏血酸允许银在抗体标记的金纳米球(10±0.5 nm)上过度生长,竞争自核生长的银原子,从而影响后者的生长动力学。利用打开的动力学窗口,以人类 IgG 为模型蛋白,实现了从 6.67 × 10-14 M(1 pg)到 3.33 × 10-9 M(50 ng)的灵敏检测。我们在此开发的非催化质子免疫吸附测定为质子 NPs 的灵敏检测增添了新的可能性。其检测斜率和分辨率比通过酶/催化剂辅助信号放大的方法更高、更好。这项工作支持了这样一种概念,即纳米粒子介导的反应灵敏度足够高,有可能在没有酶/催化剂辅助的情况下独立进行。
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A novel non-catalytic plasmonic immunoabsorbant assay amplification mechanism
The amplification of signals is dependent upon enzyme/ catalyst. Enzyme activity, specificity, pH condition, enzyme linkers, stability, storage and efficiency are some very important factors that can eventually affect the detection results. We aimed to amplify signals without the assistance of enzyme/ catalyst so that the strategy can be used in the wide range of signal detection including immune-assay. Silver nanocrystals (Ag NCs) grown from self-nucleation are used as chromogens due to their strong plasmon band at ∼ 400 nm. During 140 s of reaction time in presence of 6.10 mM CTAB and 0.61 mM of AgNO3, the 0.57 mM of Ascorbic acid allows Ag over growth on antibody-tagged gold nanospheres (10±0.5 nm) that competes silver atoms from the self-nucleation growth and thus affects the growth kinetics of the latter. By utilizing the opened kinetic window, sensitive detection has been achieved from 6.67 × 10–14 M (1 pg) to 3.33 × 10–9 M (50 ng) using human IgG as a model protein. The non-catalytic plasmonic immunoabsorbant assay we developed herein adds a new possibility of plasmonic NPs for sensitive detection. The detection slope and resolution is higher and better than those via signal amplification through the assistance of enzymes/ catalysts. This work supports the concept that nanoparticles mediated reactions are sensitive enough that they have the potential to go independently without enzymes/ catalyst assistance.
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