{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T09:45:15Z","timestamp":1791884715207,"version":"4.3.4"},"reference-count":70,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T00:00:00Z","timestamp":1791849600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T00:00:00Z","timestamp":1791849600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"abstract":"<jats:p>\n                    <jats:italic>This Viewpoint synthesises 25\u2009years of work from a single laboratory on activity\u2010based protease sensing and offers a critical appraisal of where each of the three resulting platforms stands today.<\/jats:italic>\n                    The activity of proteolytic enzymes\u2014rather than their abundance\u2014determines the outcome of inflammation, tumour progression, viral entry and barrier dysfunction. Measuring it in clinical samples is demanding: closely related proteases share substrate preferences, and biological matrices are crowded with endogenous inhibitors and competing hydrolases. Over the past 25\u2009years, our group at the University of Gdansk has developed a chemical toolkit along three complementary axes, assembled here, as a\n                    <jats:italic>probe ladder<\/jats:italic>\n                    running from solution chemistry to solid\u2010state interface. First, internally quenched chromogenic and F\u00f6rster resonance energy transfer (FRET) peptide substrates, designed by iterative deconvolution of combinatorial libraries spanning non\u2010primed and primed subsites, provide selective readouts for serine, cysteine, metallo\u2010 and viral proteases. Second, peptide\u2013quantum dot conjugates built on non\u2010toxic ZnO nanocrystals improved detection limits 20\u201330\u2010fold over the parent solution substrate in two published demonstrators. Third, peptide\u2010modified gold electrodes translate protease cleavage and molecular recognition into a label\u2010free electrochemical signal\u2014exemplified by an electrode that reads 20S proteasome activity and by an antimicrobial\u2010peptide interface for bacterial endotoxin. We applied the resulting probes in exploratory studies of bladder cancer, type\u20102 diabetes and diabetic nephropathy, and positioned them within the broader landscape of activity\u2010based protease sensing\u2014the activity\u2010based probes of the Bogyo and Cravatt laboratories, the PS\u2010SCL and HyCoSuL combinatorial approaches, and the Weissleder near\u2010infrared imaging program.\n                  <\/jats:p>","DOI":"10.1111\/febs.70729","type":"journal-article","created":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T08:52:52Z","timestamp":1791881572000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Chromogenic and\n                    <scp>FRET<\/scp>\n                    peptide substrates, quantum\u2010dot peptide conjugates and electrochemical peptide interfaces: A 25\u2010year journey from combinatorial chemistry to clinical readouts\u2014A 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