title: A competitive and reversible deactivation approach to catalysis-based quantitative assays.
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.
authors.affiliation: Department of Chemistry, University of Pittsburgh, 219 Parkman Avenue, Pittsburgh, Pennsylvania 15260, USA.
authors.forename: Kazunori
authors.initials: K
authors.lastname: Koide
authors.affiliation: Department of Chemistry, University of Pittsburgh, 219 Parkman Avenue, Pittsburgh, Pennsylvania 15260, USA.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Tracey
authors.affiliation: Process and Analytical Chemistry, Merck Research Laboratories, 126 East Lincoln Avenue, Rahway, New Jersey 07065, USA.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Bu
authors.affiliation: Process and Analytical Chemistry, Merck Research Laboratories, 126 East Lincoln Avenue, Rahway, New Jersey 07065, USA.
authors.forename: Junyong
authors.initials: J
authors.lastname: Jo
authors.affiliation: Process and Analytical Chemistry, Merck Research Laboratories, 126 East Lincoln Avenue, Rahway, New Jersey 07065, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Williams
authors.affiliation: Process and Analytical Chemistry, Merck Research Laboratories, 126 East Lincoln Avenue, Rahway, New Jersey 07065, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Welch