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Molecular Design Strategies Enabling Activity-Based Sensing and Labeling as a Platform for Probing Enzyme Activity,

Hanna I Humpel¹, Alexander P Luancing¹, Benjamin H Shostak¹

¹Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware 19716, United States.

Bioconjugate Chemistry

|February 17, 2026

Summary

Activity-based sensing and labeling (ABSL) offers sensitive detection of fleeting biological molecules. New molecular designs enhance ABSL for studying cellular signaling, enzyme activity, and reactive species.

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Area of Science:

Biochemistry
Molecular Biology
Chemical Biology

Background:

Activity-based sensing and labeling (ABSL) is a key technology for detecting transient biological analytes.
High sensitivity and spatial resolution are hallmarks of ABSL methods.
Understanding cellular signaling relies on precise detection of dynamic molecular events.

Purpose of the Study:

To highlight advanced molecular design strategies for Activity-based sensing and labeling (ABSL).
To showcase the application of ABSL in investigating critical cellular signaling pathways.
To summarize biological insights gained from ABSL of specific cellular components.

Main Methods:

Review of emerging molecular design principles for ABSL probes.
Analysis of studies employing ABSL for enzyme activity profiling.
Examination of ABSL applications for reactive oxygen and nitrogen species (ROS/RNS) detection.
Investigation of ABSL for monitoring labile transition-metal pools.

Main Results:

Novel molecular designs enable enhanced selectivity and sensitivity in ABSL.
ABSL provides significant insights into enzyme kinetics and function in situ.
ABSL successfully visualizes and quantifies dynamic changes in ROS/RNS levels.
ABSL reveals the role of labile transition metals in cellular processes.

Conclusions:

ABSL is a versatile platform for dissecting complex biological systems.
Molecular design innovations are expanding the scope and impact of ABSL.
ABSL is crucial for advancing our understanding of cellular signaling and homeostasis.