Next-Generation Enzyme-Responsive "Covalent-Assembly" Fluorescent Probes Based on Dendritic Self-Immolative Molecular Amplification
Year: 2026
Type: preprint
Abstract: Activity-based fluorogenic sensing is now regarded as a powerful approach for the accurate and sensitive detection of a myriad of important analytes, in aqueous matrices of varying complexity. Among the different molecular strategies recently proposed to devise high-performance detection schemes based on the chemical reactivity of species to be detected, the "covalent-assembly" principle is particularly promising. In order to further improve detection performances of the probes designed using this approach, we have explored their combined use with a signal amplification strategy based on the integration of a dendritic self-immolative core whose degradation should lead to in situ synthesis of several fluorophore molecules. To establish and validate this sensing system, penicillin G acylase (PGA) from E. coli was chosen as model enzyme and Pinner cyclization for in situ formation of fluorescent 2-iminocoumarin was selected as an effective biocompatible reaction. Fluorogenic AB 2 and AB 3 dendritic self-immolative molecules have been successfully synthesized. The excellent performances of PGA-responsive AB 2 dendron in terms of signal amplification and aqueous stability have been highlighted through analytical validations involving fluorescence-based enzyme assays completed with RP-HPLC-fluorescence/MS investigations. This study is a further step towards even greater popularization of the "covalent-assembly" approach in the fields of fluorogenic sensing, and bioimaging.
Source: ChemRxiv
Institutions: Université de Bourgogne, Institut de Chimie Moléculaire de l'Université de Bourgogne, Université Bourgogne Europe, University of Chemistry and Technology, Prague, University of Namur, Université Paris Sciences et Lettres, Université Paris-Saclay, Namur Research Institute for Life Sciences, Institut Curie, Université Côte d'Azur, Institut de Chimie de Nice
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: green
Awards: A luminogenic real-time detection system for monitoring enzymatic manufacturing of oligonucleotides, G1918669190, G2841865453, G3527965702, G3535982825, G3562746947, Butyrylcholinesterase-based fluorescent and/or colorimetric detection of organophosphorus nerve agents, G4656594441, G8800819381, G880779785
OpenAlex ID: W7205016387