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Single-Molecule Activity Profiling of Glycosidase Proteoforms Using Water-Soluble Fluorogenic Probes

Kaneko, A.; Fujita, K.; Uchida, T.; Mizuno, T.; Iwasaka, T.; Kusuhara, H.; Urano, Y.; Komatsu, T.

2026-05-24 biochemistry
10.64898/2026.05.22.727124 bioRxiv
Show abstract

In this study, we report a series of highly water-soluble fluorogenic probes for glycosidases useful for microdevice-based single-molecule enzyme activity profiling (SEAP). The assay was able to detect trace glycosidase activities in blood with high sensitivity and with proteoform resolution. This platform revealed the potential of -mannosidase as sensitive blood-based liquid-biopsy biomarkers for liver injury, highlighting the potential of this approach for activity-based diagnostics.

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