A competitive and reversible deactivation approach to catalysis-based quantitative assays

Kazunori Koide (), Matthew P. Tracey, Xiaodong Bu, Junyong Jo, Michael J. Williams and Christopher J. Welch ()
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Kazunori Koide: University of Pittsburgh
Matthew P. Tracey: University of Pittsburgh
Xiaodong Bu: Process and Analytical Chemistry, Merck Research Laboratories
Junyong Jo: Process and Analytical Chemistry, Merck Research Laboratories
Michael J. Williams: Process and Analytical Chemistry, Merck Research Laboratories
Christopher J. Welch: Process and Analytical Chemistry, Merck Research Laboratories

Nature Communications, 2016, vol. 7, issue 1, 1-7

Abstract: Abstract 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.

Date: 2016
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DOI: 10.1038/ncomms10691

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