A competitive and reversible deactivation approach to catalysis-based quantitative assays.
A competitive and reversible deactivation approach to catalysis-based quantitative assays.
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基于催化的定量测定的竞争性和可逆失活方法。
DOI：
10.1038/ncomms10691
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发表时间：
2016-02-19
影响因子：
16.6
通讯作者：
Welch CJ
中科院分区：
文献类型：
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作者：
Koide K;Tracey MP;Bu X;Jo J;Williams MJ;Welch CJ
基于催化的信号放大使得光学分析具有高度灵敏度，并且在化学和生物化学研究中具有广泛的用途。然而，必须对测定进行微调以避免信号饱和、底物耗尽和非线性性能。此外，一旦停止，这样的测定不能重新开始，相对于分析物浓度将动态范围限制在两个数量级。此外，由于快速信号饱和，在催化条件下难以定量丰富的分析物。在此，我们报告了一种方法，其中催化反应与伴随的催化剂失活或信号产生所需的试剂消耗竞争。因此，信号生成在有限的时间内进行，然后自主且可逆地停止。在两个基于催化的测定中，我们展示了重新启动自主停滞的反应，能够准确测量超过五个数量级，包括高于底物浓度的分析物水平。这表明基于催化的测定的动态范围可以通过竞争性和可逆失活而显著拓宽。 催化体系的分析，特别是那些简单的比色读数，是有用的快速评估性能。在这里，作者报告了一种基于在宽范围内工作的同时显色反应的测定，该反应可以停止以允许测量，随后重新开始。
Catalysis-based signal amplification makes optical assays highly sensitive and widely useful in chemical and biochemical research. However, assays must be fine-tuned to avoid signal saturation, substrate depletion and nonlinear performance. Furthermore, once stopped, such assays cannot be restarted, limiting the dynamic range to two orders of magnitude with respect to analyte concentrations. In addition, abundant analytes are difficult to quantify under catalytic conditions due to rapid signal saturation. Herein, we report an approach in which a catalytic reaction competes with a concomitant inactivation of the catalyst or consumption of a reagent required for signal generation. As such, signal generation proceeds for a limited time, then autonomously and reversibly stalls. In two catalysis-based assays, we demonstrate restarting autonomously stalled reactions, enabling accurate measurement over five orders of magnitude, including analyte levels above substrate concentration. This indicates that the dynamic range of catalysis-based assays can be significantly broadened through competitive and reversible deactivation. Assays for catalytic systems—particularly ones with simple colorimetric readouts—are useful for the rapid evaluation of performance. Here, the authors report an assay based on a concurrent colour-forming reaction working across a wide range that can be stopped to allow measurements and subsequently restarted.