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In this work, we developed an ECL biosensor capable of detecting microcystin-LR (MC-LR) at trace levels. The system combines three-dimensional pyrene-based covalent organic frameworks (3D Py-COF) with an entropy-driven DNA cyclic amplification process, exhibiting excellent sensitivity and reliability. The unique 3D topological architecture of the Py-COF spatially isolates the pyrene nodes, effectively suppressing intermolecular \u03c0\u2013\u03c0 stacking and mitigating the ACQ effect. Furthermore, in situ-loaded silver nanoparticles (Ag NPs) serve as potent internal catalysts, facilitating potassium persulfate (K2S2O8) transformation to generate sulfate radicals (SO4\u2022\u2013). The design not only significantly enhances ECL emission but also mitigates electrode passivation effects. By combining the 3D Py-COF with a designed entropy-driven DNA cyclic amplification strategy, the ECL detection system achieves signal transduction and amplification driven by the synergistic action of aptamer recognition and endonuclease cleavage. The proposed biosensor greatly improved the reliability of detection, demonstrating a broad linear range spanning from 1.0 pg\/mL to 100 ng\/mL, with a limit of detection as low as 0.51 pg\/mL. Taking advantage of strong selectivity and dependable reproducibility, the present work introduces an innovative method for precise detection of trace toxins in water systems.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c04377","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T06:36:26Z","timestamp":1788330986000},"page":"27137-27145","source":"Crossref","is-referenced-by-count":0,"title":["Silver Nanoparticle-Loaded\n3D Pyrene-Based Covalent\nOrganic Frameworks Combined with an Entropy-Driven Amplification Strategy\nfor Microcystin-LR Detection"],"prefix":"10.1021","volume":"98","author":[{"given":"Qingfa","family":"Li","sequence":"first","affiliation":[{"name":"University of Jinan , , ,","place":["Jinan, P. R. China, 250022"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujie","family":"Han","sequence":"additional","affiliation":[{"name":"University of Jinan , , ,","place":["Jinan, P. R. China, 250022"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuangzhuang","family":"Ru","sequence":"additional","affiliation":[{"name":"University of Jinan , , ,","place":["Jinan, P. R. China, 250022"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4321-4282","authenticated-orcid":true,"given":"Xiang","family":"Ren","sequence":"additional","affiliation":[{"name":"University of Jinan , , ,","place":["Jinan, P. R. 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China, 250022"]},{"name":"Sungkyunkwan University , , ,","place":["Suwon, Republic of Korea, 16419"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,28]]},"reference":[{"key":"2026091511083016200_cit1","doi-asserted-by":"publisher","first-page":"116413","DOI":"10.1016\/j.bios.2024.116413","article-title":"Aptameric Photonic Structure-Based\nOptical Biosensor for the Detection of Microcystin","volume":"260","author":"Hussain","year":"2024","journal-title":"Biosens. Bioelectron."},{"issue":"37","key":"2026091511083016200_cit2","doi-asserted-by":"publisher","first-page":"15050","DOI":"10.1021\/acs.analchem.4c03495","article-title":"Aggregation-Induced\nElectrochemiluminescence of Ir(ppy)3-Functionalized ZIF-8\nfor Microcystin-LR Detection Via the Trans-Cleavage Activity of CRISPR-Cas12a","volume":"96","author":"Shi","year":"2024","journal-title":"Anal. 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Eng.\nJ."},{"issue":"22","key":"2026091511083016200_cit36","doi-asserted-by":"publisher","first-page":"8679","DOI":"10.1021\/acs.analchem.3c01285","article-title":"Sulfur Quantum\nDot-Based Electrochemiluminescence Resonance\nEnergy Transfer System for a Dual-Mode Ultrasensitive Detection of\nMC-LR Using Ag+ as a Signal Amplification Switch","volume":"95","author":"Han","year":"2023","journal-title":"Anal. 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