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This study introduces tandem assays, dynamic host-dye systems that monitor analyte concentrations altered by biological processes. These chemosensors offer real-time insights into biochemical pathways for pharmaceutical and environmental applications.

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Area of Science:

  • Supramolecular Chemistry
  • Analytical Chemistry

Background:

  • Host-guest chemistry, pioneered by Pedersen's crown ether synthesis, has advanced significantly in designing selective receptors for analyte sensing.
  • Traditional chemosensors combine macrocyclic receptors with indicator dyes, often using indicator displacement assays, primarily for static analyte concentration determination.
  • The application of these assays in dynamic biological systems, such as membranes and enzymes, remains less explored.

Purpose of the Study:

  • To introduce and explore the principles of tandem assays, a novel class of dynamically analyte-responsive host-dye systems.
  • To demonstrate the potential of tandem assays for real-time monitoring of analyte concentration changes induced by biological reactions.
  • To highlight the applicability of these assays in biochemical research, pharmaceutical development, and environmental monitoring.

Main Methods:

  • Development of supramolecular tandem assays, including product- and substrate-selective, domino, and enzyme-coupled variants.
  • Design of tandem membrane assays for monitoring analyte translocation across membranes.
  • Utilizing macrocycle-fluorescent dye complexes to generate fluorescence responses (switch-on or switch-off) indicative of analyte concentration changes.

Main Results:

  • Demonstrated various tandem assay configurations, including enzyme-coupled assays for enzymes like choline oxidase and acetylcholineesterase, and protease activity monitoring.
  • Successfully implemented tandem membrane assays for tracking analyte translocation mediated by channel proteins.
  • Showcased proof-of-principle for environmental monitoring with sensing ensembles for volatile hydrocarbons.

Conclusions:

  • Tandem assays represent a significant advancement in chemosensing, enabling dynamic monitoring of analyte concentrations influenced by biological processes.
  • These assays offer valuable tools for real-time analysis in biochemical and pharmaceutical research, as well as for environmental monitoring.
  • The versatility of tandem assays, including membrane and enzyme-coupled variants, expands their utility across diverse scientific disciplines.