{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:43:45Z","timestamp":1792806225385,"version":"4.5.0"},"reference-count":55,"publisher":"American Chemical Society (ACS)","issue":"11","license":[{"start":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T00:00:00Z","timestamp":1761091200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T00:00:00Z","timestamp":1761091200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T00:00:00Z","timestamp":1761091200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100005031","name":"Bhabha Atomic Research Centre","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/501100005031","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,11,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Accurate detection of biomolecules such as protamine and trypsin is critical due to their significant clinical and diagnostic roles. Protamine sulfate, a highly cationic peptide, is routinely used to reverse the anticoagulant effects of heparin, necessitating precise monitoring to avoid adverse outcomes. Trypsin, a serine protease, is essential in digestion and implicated in disorders such as pancreatitis, cystic fibrosis, and various cancers, making it a valuable disease biomarker. In this work, we introduce a fluorescence-based sensing platform utilizing commercially available carboxylated tetraphenylethylene (CTPE), an aggregation-induced emission (AIE) luminogen. This is a unique demonstration of dual protamine\/trypsin detection using a readily available AIE luminogen, eliminating the need for custom synthesis and simplifying real-world deployment. CTPE forms highly emissive aggregates in aqueous buffer via electrostatic complexation with protamine, leading to significant fluorescence enhancement due to restricted intramolecular motion. This enables highly sensitive protamine detection with a detection limit of 0.7 nM, demonstrating successful application in diluted human serum. Comprehensive photophysical analyses\u2500including absorption, steady-state and time-resolved fluorescence spectroscopy\u2500confirmed the formation and disassembly dynamics of the CTPE\u2013PrS complex. Upon trypsin-mediated enzymatic cleavage of protamine, the aggregates dissociate, triggering measurable fluorescence quenching. This system achieves trypsin detection down to 11.8 pM, maintaining a linear response from 0 to 20 nM, even in 20% human urine, underscoring its robustness in complex biological matrices. This dual-analyte sensing platform, based on a readily available AIE fluorophore, offers a sensitive, selective, and scalable approach for real-sample analysis and enzyme activity monitoring.<\/jats:p>","DOI":"10.1021\/acsabm.5c01343","type":"journal-article","created":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T11:30:17Z","timestamp":1761132617000},"page":"9993-10005","source":"Crossref","is-referenced-by-count":7,"title":["A Dual-Detection AIE-Based Fluorescent Probe for Sensitive\nDetection of Protamine and Trypsin in Biological Media"],"prefix":"10.1021","volume":"8","author":[{"given":"Shreya Y.","family":"Garge","sequence":"first","affiliation":[{"name":"Bhabha Atomic Research Centre , , ,","place":["Mumbai, India, 400085"]},{"name":"SVKM\u2019s Dr. Bhanuben Nanavati College of Pharmacy, Vile Parle , ,","place":["Mumbai, India, 400056"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vasanti","family":"Suvarna","sequence":"additional","affiliation":[{"name":"SVKM\u2019s Dr. Bhanuben Nanavati College of Pharmacy, Vile Parle , ,","place":["Mumbai, India, 400056"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shraddha V.","family":"Sonavane","sequence":"additional","affiliation":[{"name":"Homi Bhabha National Institute , Training School complex, Anushaktinagar , ,","place":["Mumbai, India, 400094"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4612-547X","authenticated-orcid":true,"given":"Prabhat K.","family":"Singh","sequence":"additional","affiliation":[{"name":"Bhabha Atomic Research Centre , , ,","place":["Mumbai, India, 400085"]},{"name":"Homi Bhabha National Institute , Training School complex, Anushaktinagar , ,","place":["Mumbai, India, 400094"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2025,10,22]]},"reference":[{"issue":"5","key":"2026072914063152400_cit1","first-page":"659","article-title":"The heparin-protamine\ninteraction","volume":"40","author":"Carr","year":"1999","journal-title":"A review. 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Thromb.\/Hemost."},{"issue":"7","key":"2026072914063152400_cit5","doi-asserted-by":"publisher","first-page":"624","DOI":"10.1097\/EJA.0b013e32833731bd","article-title":"The effects of protamine\noverdose on coagulation parameters as measured by the thrombelastograph","volume":"27","author":"Khan","year":"2010","journal-title":"Eur. J. Anaesthesiol."},{"issue":"6","key":"2026072914063152400_cit6","doi-asserted-by":"publisher","first-page":"877","DOI":"10.3390\/pharmaceutics13060877","article-title":"Protamine-Based Strategies for RNA\nTransfection","volume":"13","author":"Jarzebska","year":"2021","journal-title":"Pharmaceutics."},{"key":"2026072914063152400_cit7","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1016\/j.ejpb.2015.09.019","article-title":"Protamine-based\nnanoparticles as new antigen delivery systems","volume":"97","author":"Gonza\u0301lez-Aramundiz","year":"2015","journal-title":"Eur. J. Pharm. Biopharm"},{"issue":"6","key":"2026072914063152400_cit8","doi-asserted-by":"publisher","first-page":"1508","DOI":"10.3390\/nano11061508","article-title":"Use of Protamine in\nNanopharmaceuticals\u2500A Review","volume":"11","author":"Ruseska","year":"2021","journal-title":"Nanomater."},{"issue":"3","key":"2026072914063152400_cit9","doi-asserted-by":"publisher","first-page":"314","DOI":"10.1007\/BF01318124","article-title":"Activation\nof pancreatic zymogens","volume":"31","author":"Rinderknecht","year":"1986","journal-title":"Dig. Dis. 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Sci."},{"key":"2026072914063152400_cit30","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccr.2021.214135","article-title":"AIE-featured tetraphenylethylene nanoarchitectures in biomedical\napplication: Bioimaging, drug delivery and disease treatment","volume":"447","author":"Sharath Kumar","year":"2021","journal-title":"Coord. Chem. Rev."},{"issue":"5","key":"2026072914063152400_cit31","doi-asserted-by":"publisher","first-page":"1790","DOI":"10.1016\/j.cclet.2020.12.031","article-title":"Aggregation-induced emission or aggregation-caused\nquenching? Impact\nof covalent bridge between tetraphenylethene and naphthalimide","volume":"32","author":"Ma","year":"2021","journal-title":"Chin. Chem. Lett."},{"key":"2026072914063152400_cit32","doi-asserted-by":"publisher","DOI":"10.1016\/j.snb.2021.130517","article-title":"An AIEgen\u2013protamine\nassembly\/disassembly based\nfluorescence turn-on probe for sensing alkaline phosphatase","volume":"346","author":"Kaur","year":"2021","journal-title":"Sens. Actuators B Chem."},{"key":"2026072914063152400_cit33","doi-asserted-by":"publisher","DOI":"10.1016\/j.snb.2021.131016","article-title":"A tetracationic\naggregation induced emission-based probe for efficient and improved\ndetection of Heparin","volume":"353","author":"Pandey","year":"2022","journal-title":"Sens. Actuators B: Chem."},{"issue":"41","key":"2026072914063152400_cit34","doi-asserted-by":"publisher","first-page":"8414","DOI":"10.1039\/D0OB01661F","article-title":"An ATP responsive\nfluorescent supramolecular assembly based on a polyelectrolyte and\nan AIE active tetraphenylethylene derivative","volume":"18","author":"Singh","year":"2020","journal-title":"Org. Biomol. Chem."},{"issue":"20","key":"2026072914063152400_cit35","doi-asserted-by":"publisher","first-page":"18344","DOI":"10.1021\/am505791f","article-title":"Amino-Modified\nTetraphenylethene Derivatives as Nucleic Acid Stain: Relationship\nbetween the Structure and Sensitivity","volume":"6","author":"Xu","year":"2014","journal-title":"ACS Appl.\nMater. Interfaces"},{"issue":"12","key":"2026072914063152400_cit36","doi-asserted-by":"publisher","first-page":"3209","DOI":"10.1039\/D0MH01303J","article-title":"The largest\nCPL enhancement by further assembly of\nself-assembled superhelices based on the helical TPE macrocycle","volume":"7","author":"Yuan","year":"2020","journal-title":"Mater. Horiz."},{"issue":"5","key":"2026072914063152400_cit37","doi-asserted-by":"publisher","DOI":"10.1016\/j.cclet.2024.110724","article-title":"Significantly boosting\ncircularly polarized luminescence by synergy\nof helical and planar chirality","volume":"36","author":"Ye","year":"2025","journal-title":"Chin. Chem.\nLett."},{"issue":"29","key":"2026072914063152400_cit38","doi-asserted-by":"publisher","first-page":"3586","DOI":"10.1039\/D3AY00753G","article-title":"Self-assembled\nsuper-small AIEgen nanoprobe for highly sensitive and selective detection\nof protamine and trypsin","volume":"15","author":"Zhang","year":"2023","journal-title":"Anal. Methods."},{"issue":"15","key":"2026072914063152400_cit39","doi-asserted-by":"publisher","first-page":"3786","DOI":"10.1039\/D4TB00264D","article-title":"Expanding the scope\nof self-assembled supramolecular\nbiosensors: a highly selective and sensitive enzyme-responsive AIE-based\nfluorescent biosensor for trypsin detection and inhibitor screening","volume":"12","author":"Kaur","year":"2024","journal-title":"J. Mater. Chem. 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Liq."},{"issue":"25","key":"2026072914063152400_cit48","doi-asserted-by":"publisher","first-page":"10429","DOI":"10.1021\/acs.jpcc.2c01423","article-title":"Aggregation-Induced\nEmission in Phenothiazine-Based Fluorophores: An Insight into the\nExcited State and Aggregate Formation Mechanism","volume":"126","author":"Cesaretti","year":"2022","journal-title":"J. Phys. Chem. C"},{"issue":"6","key":"2026072914063152400_cit49","doi-asserted-by":"publisher","first-page":"608","DOI":"10.1074\/mcp.T400003-MCP200","article-title":"Trypsin Cleaves\nExclusively C-terminal\nto Arginine and Lysine Residues","volume":"3","author":"Olsen","year":"2004","journal-title":"Mol. Cell.\nProteomics."},{"issue":"45","key":"2026072914063152400_cit50","doi-asserted-by":"publisher","first-page":"26765","DOI":"10.1039\/D0RA03970E","article-title":"A simple fluorescence\nassay for trypsin\nthrough a protamine-induced carbon quantum dot-quenching aggregation\nplatform","volume":"10","author":"Chen","year":"2020","journal-title":"RSC Adv."},{"key":"2026072914063152400_cit51","doi-asserted-by":"publisher","DOI":"10.1016\/j.molliq.2021.115980","article-title":"An anionic tetraphenyl\nethylene based simple and rapid fluorescent probe for detection of\ntrypsin and paraoxon methyl","volume":"333","author":"Kaur","year":"2021","journal-title":"J. Mol. Liq."},{"issue":"5","key":"2026072914063152400_cit52","doi-asserted-by":"publisher","first-page":"2802","DOI":"10.1039\/C5SC00524H","article-title":"Molecular\nglues for manipulating\nenzymes: trypsin inhibition by benzamidine-conjugated molecular glues","volume":"6","author":"Mogaki","year":"2015","journal-title":"Chem. 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Gastroenterol."},{"issue":"1","key":"2026072914063152400_cit55","doi-asserted-by":"publisher","first-page":"340","DOI":"10.1039\/C4AN01731E","article-title":"Quantitative serine\nprotease assays based on formation\nof copper(II)\u2013oligopeptide complexes","volume":"140","author":"Ding","year":"2015","journal-title":"Analyst."}],"container-title":["ACS Applied Bio Materials"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsabm.5c01343","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/aabmcb\/article-pdf\/8\/11\/9993\/66301828\/acsabm.5c01343.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/aabmcb\/article-pdf\/8\/11\/9993\/66301828\/acsabm.5c01343.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T18:13:46Z","timestamp":1785348826000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/aabmcb\/article\/8\/11\/9993\/3751537\/A-Dual-Detection-AIE-Based-Fluorescent-Probe-for"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,22]]},"references-count":55,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2025,10,22]]},"published-print":{"date-parts":[[2025,11,17]]}},"URL":"https:\/\/doi.org\/10.1021\/acsabm.5c01343","relation":{},"ISSN":["2576-6422"],"issn-type":[{"value":"2576-6422","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,22]]}}}