{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T22:42:08Z","timestamp":1785796928682,"version":"3.56.0"},"reference-count":39,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:00:00Z","timestamp":1764288000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>The widespread use of organophosphorus pesticides (OPs) in Chinese herbal medicines cultivation raises urgent concerns about residue contamination. Conventional detection methods [e.g., gas chromatography-mass spectrometry (GC-MS) and enzyme-linked immunosorbent assay (ELISA)] suffer from poor portability and instability of antibody inactivation in complex matrices, hindering on-site analysis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>This study proposed a novel \u201celectrostatic adsorption-driven cascade reaction chain\u201d strategy for rapid detection of acetylcholinesterase (AChE) activity and OPs. Leveraging the electrostatic self-assembly between a positively charged acetylcholine chloride (ACh, 26.47 \u00b1 1.63 mV) and a negatively charged choline oxidase (CHO, \u221230.81 \u00b1 1.85 mV), a nanoscale fluorescence sensor (CA-B NPs) was constructed by encapsulating the Azo-Bodipy 685. This design created a spatially confined and componentially co-localized nanoreactor that restricted substrate diffusion distance to the nanoscale and utilized a dual-enzyme cascade system (AChE-CHO) to yield a signal amplification effect.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The obtained CA-B NPs exhibited excellent analytical performance, including: (1) a low detection limit of 4.1 ng\/mL for triazophos; (2) high recovery of 88.13%\u2212113.09% in complex Citrus reticulata Blanco matrices, along with strong anti-interference capabilities by organically dividing the reaction and detection sections; (3) a total assay time of only 20 min for real samples, suitable for rapid, on-site, high-throughput screening.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      This study not only embedded the entire reaction chain [AChE-CHO-hydrogen peroxide (H\n                      <jats:sub>2<\/jats:sub>\n                      O\n                      <jats:sub>2<\/jats:sub>\n                      )] into the sensor to improve space utilization efficiency and detection efficiency, but also established a novel paradigm for enzyme spatial organization based on electrostatic complementarity, providing new insights into the rational design of nanostructured multi-enzyme sensing platforms.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fphar.2025.1679948","type":"journal-article","created":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T06:28:08Z","timestamp":1764311288000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Electrostatically driven fluorescent sensor for rapid detection of AChE activity and organophosphate pesticides via dual-enzyme cascade amplification"],"prefix":"10.3389","volume":"16","author":[{"given":"Xiaoyi","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kunhui","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sitong","family":"Lai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guojing","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meifang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuhong","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lan","family":"Ma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xie-An","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Lei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bing","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,11,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"386","DOI":"10.3390\/bios13030386","article-title":"An amperometric acetylcholine biosensor based on Co-Immobilization of enzyme nanoparticles onto nanocomposite","volume":"13","author":"Ahlawat","year":"2023","journal-title":"Biosensors"},{"key":"B2","doi-asserted-by":"publisher","first-page":"2419499","DOI":"10.1002\/adfm.202419499","article-title":"Enzyme-nanozyme Cascade flow reactor synergy with deep learning for differentiation and point-of-care testing of multiple organophosphorus pesticides","volume":"35","author":"Bai","year":"2025","journal-title":"Adv. Funct. Mat."},{"key":"B3","doi-asserted-by":"publisher","first-page":"3321","DOI":"10.3390\/ijms23063321","article-title":"Fluorescence-based binding characterization of small molecule ligands targeting CUG RNA repeats","volume":"23","author":"Chang","year":"2022","journal-title":"Int. J. Mol. Sci."},{"key":"B4","doi-asserted-by":"publisher","first-page":"5917","DOI":"10.1021\/acsomega.7b00863","article-title":"Multiresidue method for the rapid determination of pesticide residues in tea using ultra performance liquid chromatography orbitrap high resolution mass spectrometry and In-Syringe dispersive solid phase extraction","volume":"2","author":"Chen","year":"2017","journal-title":"ACS Omega"},{"key":"B5","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1016\/j.cjac.2021.09.004","article-title":"On-line immobilized acetylcholinesterase microreactor based on capillary electrophoresis for the determination of organophosphorus pesticide residues","volume":"49","author":"Chen","year":"2021","journal-title":"Chin. J. Anal. Chem."},{"key":"B6","doi-asserted-by":"publisher","first-page":"647","DOI":"10.1080\/09540105.2019.1612331","article-title":"A sensitive lateral flow immunoassay for the multiple residues of five adamantanes","volume":"30","author":"Gao","year":"2019","journal-title":"Food Agric. Immunol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"p158","DOI":"10.1097\/HM9.0000000000000071","article-title":"Version updates of strategies for drug discovery based on effective constituents of traditional Chinese medicine","volume":"3","author":"Ge","year":"2023","journal-title":"Acupunct. Herb. Med."},{"key":"B8","doi-asserted-by":"publisher","first-page":"1073","DOI":"10.1515\/pac-2019-0204","article-title":"Carbohydrates and BODIPYs: access to bioconjugatable and water-soluble BODIPYs","volume":"91","author":"Gomez","year":"2019","journal-title":"Pure Appl. Chem."},{"key":"B9","doi-asserted-by":"publisher","first-page":"123145","DOI":"10.1016\/j.talanta.2021.123145","article-title":"Recent advances and perspectives of enzyme-based optical biosensing for organophosphorus pesticides detection","volume":"240","author":"Gong","year":"2022","journal-title":"Talanta"},{"key":"B10","doi-asserted-by":"publisher","first-page":"11244","DOI":"10.1021\/acs.jafc.9b03624","article-title":"One-step facile synthesis of nitrogen-doped carbon dots: a ratiometric fluorescent probe for evaluation of acetylcholinesterase activity and detection of organophosphorus pesticides in tap water and food","volume":"67","author":"Huang","year":"2019","journal-title":"J. Agric."},{"key":"B11","doi-asserted-by":"publisher","first-page":"24190","DOI":"10.1039\/d0ra03055d","article-title":"CdTe QD-based inhibition and reactivation assay of acetylcholinesterase for the detection of organophosphorus pesticides","volume":"10","author":"Korram","year":"2020","journal-title":"RSC Adv."},{"key":"B12","doi-asserted-by":"publisher","first-page":"64769","DOI":"10.1039\/c6ra13185a","article-title":"Acetylcholinesterase (AChE)-mediated immobilization of silver nanoparticles for the detection of organophosphorus pesticides","volume":"6","author":"Kumar","year":"2016","journal-title":"RSC Adv."},{"key":"B13","doi-asserted-by":"publisher","first-page":"344625","DOI":"10.1016\/j.aca.2025.344625","article-title":"Pd-modified MXene enabled electrochemical sensing platform for ratiometric detection of acetylcholinesterase inhibitors via modulation effect of Prussian blue","volume":"1376","author":"Li","year":"2025","journal-title":"Anal. Chim. Acta."},{"key":"B14","doi-asserted-by":"publisher","first-page":"1265","DOI":"10.6023\/cjoc202111001","article-title":"Progress in the synthesis of boron dipyrromethene (BODIPY) fluorescent dyes","volume":"42","author":"Liu","year":"2022","journal-title":"Chin. J. Org. Chem."},{"key":"B15","doi-asserted-by":"publisher","first-page":"9186536","DOI":"10.1155\/2022\/9186536","article-title":"Bacterial metabolomics: sample preparation methods","volume":"2022","author":"Mohd Kamal","year":"2022","journal-title":"Biochem. Research International"},{"key":"B16","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1016\/j.snb.2018.03.181","article-title":"Gold nanoprobe for inhibition and reactivation of acetylcholinesterase: an application to detection of organophosphorus pesticides","volume":"267","author":"Satnami","year":"2018","journal-title":"Sens. Actuators B Chem."},{"key":"B17","doi-asserted-by":"publisher","first-page":"899","DOI":"10.3390\/foods11070899","article-title":"Simultaneous determination of pyridate, quizalofop-Ethyl and cyhalofop-butyl residues in agricultural products using liquid chromatography-tandem mass spectrometry","volume":"11","author":"Shim","year":"2022","journal-title":"Foods Basel, Switz."},{"key":"B18","doi-asserted-by":"publisher","first-page":"1388","DOI":"10.3390\/molecules22091388","article-title":"Determination of coenzyme A and acetyl-coenzyme A in biological samples using HPLC with UV detection","volume":"22","author":"Shurubor","year":"2017","journal-title":"Mol. Basel, Switz."},{"key":"B19","doi-asserted-by":"publisher","first-page":"5563759","DOI":"10.1155\/2022\/5563759","article-title":"A common feature of pesticides: oxidative stress-the role of oxidative stress in pesticide-induced toxicity","volume":"2022","author":"Sule","year":"2022","journal-title":"Oxid. Med. Cell. Longev."},{"key":"B20","doi-asserted-by":"publisher","first-page":"9999","DOI":"10.3390\/ijms14059999","article-title":"A novel colorimetric immunoassay utilizing the peroxidase mimicking activity of magnetic nanoparticles","volume":"14","author":"Woo","year":"2013","journal-title":"Int. J. Mol. Sci."},{"key":"B21","doi-asserted-by":"publisher","first-page":"123619","DOI":"10.1016\/j.jhazmat.2020.123619","article-title":"Double-enzymes-mediated Fe2+\/Fe3+ conversion as magnetic relaxation switch for pesticide residues sensing","volume":"403","author":"Wu","year":"2021","journal-title":"J. Hazard. Mat."},{"key":"B22","doi-asserted-by":"publisher","first-page":"115341","DOI":"10.1016\/j.bios.2023.115341","article-title":"Near-infrared-excitable acetylcholinesterase-activated fluorescent probe for sensitive and anti-interference detection of pesticides in colored food","volume":"233","author":"Wu","year":"2023","journal-title":"Biosens. Bioelectron."},{"key":"B23","doi-asserted-by":"publisher","first-page":"121362","DOI":"10.1016\/j.talanta.2020.121362","article-title":"A thiamine-triggered fluormetric assay for acetylcholinesterase activity and inhibitor screening based on oxidase-like activity of MnO2 nanosheets","volume":"221","author":"Xiao","year":"2021","journal-title":"Talanta"},{"key":"B24","doi-asserted-by":"publisher","first-page":"114147","DOI":"10.1016\/j.microc.2025.114147","article-title":"Fluorescent method for acetylcholinesterase activity detection and its inhibitors screening","volume":"214","author":"Xiao","year":"2025","journal-title":"Microchem. J."},{"key":"B25","doi-asserted-by":"publisher","first-page":"637","DOI":"10.1016\/j.jpha.2022.05.004","article-title":"A highly sensitive bio-barcode immunoassay for multi-residue detection of organophosphate pesticides based on fluorescence anti-quenching","volume":"12","author":"Xu","year":"2022","journal-title":"J. Pharm. Anal."},{"key":"B26","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1007\/s00216-022-04400-0","article-title":"Engineering an Ag\/Au bimetallic nanoparticle-based acetylcholinesterase SERS biosensor for in situ sensitive detection of organophosphorus pesticide residues in food","volume":"415","author":"Xu","year":"2023","journal-title":"Anal. Bioanal. Chem."},{"key":"B27","doi-asserted-by":"publisher","first-page":"145483","DOI":"10.1016\/j.cej.2023.145483","article-title":"Bismuth atom-doped gold aerogels for the detection of acetylcholinesterase activity and organophosphorus inhibitor","volume":"474","author":"Yan","year":"2023","journal-title":"Chem. Eng. J."},{"key":"B28","doi-asserted-by":"publisher","first-page":"12404","DOI":"10.1021\/acs.analchem.4c01901","article-title":"Monitoring enzymatic reaction kinetics and activity assays in confined nanospace","volume":"96","author":"Yang","year":"2024","journal-title":"Anal. Chem."},{"key":"B29","doi-asserted-by":"publisher","first-page":"122","DOI":"10.1016\/j.aca.2016.06.029","article-title":"In situ hydrothermal growth of ZnO\/g-C3N4 nanoflowers coated solid-phase microextraction fibers coupled with GC-MS for determination of pesticides residues","volume":"934","author":"Zhang","year":"2016","journal-title":"Anal. Chim. Acta."},{"key":"B30","doi-asserted-by":"publisher","first-page":"p131","DOI":"10.1097\/HM9.0000000000000162","article-title":"Ten recommendations for the high-quality development of traditional Chinese medicine in the new era","volume":"5","author":"Zhang","year":"","journal-title":"Acupunct. Herb. Med."},{"key":"B31","doi-asserted-by":"publisher","first-page":"127020","DOI":"10.1016\/j.talanta.2024.127020","article-title":"Palladium-platinum bimetallic modified MXene nanozyme for highly sensitive detection of active substances with acetylcholinesterase inhibitory effect","volume":"282","author":"Zhang","year":"","journal-title":"Talanta"},{"key":"B32","doi-asserted-by":"publisher","first-page":"11658","DOI":"10.1021\/acs.analchem.8b03250","article-title":"Metallic transition metal dichalcogenide nanosheets as an effective and biocompatible transducer for electrochemical detection of pesticide","volume":"90","author":"Zhao","year":"2018","journal-title":"Anal. Chem."},{"key":"B33","doi-asserted-by":"publisher","first-page":"121358","DOI":"10.1016\/j.jhazmat.2019.121358","article-title":"Self-reduction bimetallic nanoparticles on ultrathin MXene nanosheets as functional platform for pesticide sensing","volume":"384","author":"Zhao","year":"","journal-title":"J. Hazard. Mat."},{"key":"B34","doi-asserted-by":"publisher","first-page":"112636","DOI":"10.1016\/j.bios.2020.112636","article-title":"Smart plant-wearable biosensor for in-situ pesticide analysis","volume":"170","author":"Zhao","year":"","journal-title":"Biosens. Bioelectron."},{"key":"B35","doi-asserted-by":"publisher","first-page":"117152","DOI":"10.1016\/j.trac.2023.117152","article-title":"Nanozyme-based biosensor for organophosphorus pesticide monitoring: functional design, biosensing strategy and detection application","volume":"165","author":"Zhao","year":"","journal-title":"Trends Anal. Chem."},{"key":"B36","doi-asserted-by":"publisher","first-page":"4503","DOI":"10.1021\/acs.analchem.2c05313","article-title":"Ultrasensitivity detecting AChE through \u201cCovalent Assembly\u201d and signal amplification strategic approaches and applied to screen its inhibitor","volume":"95","author":"Zhao","year":"","journal-title":"Anal. Chem."},{"key":"B37","doi-asserted-by":"publisher","first-page":"p171","DOI":"10.1097\/HM9.0000000000000119","article-title":"Safety evaluation of traditional Chinese medicine: new era, new strategy. Acupunct. Herb","volume":"4","author":"Zhao","year":"2024","journal-title":"Med"},{"key":"B38","doi-asserted-by":"publisher","first-page":"1112","DOI":"10.3390\/microorganisms10061112","article-title":"A novel organophosphorus acid anhydrolase from deep sea sediment with high degradation efficiency for organophosphorus pesticides and nerve agent","volume":"10","author":"Zheng","year":"2022","journal-title":"Microorganisms"},{"key":"B39","doi-asserted-by":"publisher","first-page":"84","DOI":"10.3390\/bios12020084","article-title":"Development of a rapid gold nanoparticle immunochromatographic strip based on the nanobody for detecting 2,4-DichloRophenoxyacetic acid","volume":"12","author":"Zhou","year":"2022","journal-title":"Biosensors"}],"container-title":["Frontiers in Pharmacology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2025.1679948\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T06:28:08Z","timestamp":1764311288000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2025.1679948\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,28]]},"references-count":39,"alternative-id":["10.3389\/fphar.2025.1679948"],"URL":"https:\/\/doi.org\/10.3389\/fphar.2025.1679948","relation":{},"ISSN":["1663-9812"],"issn-type":[{"value":"1663-9812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,28]]},"article-number":"1679948"}}