{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T14:06:34Z","timestamp":1789567594190,"version":"build-2803163510"},"reference-count":39,"publisher":"American Chemical Society (ACS)","issue":"36","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22361132536"],"award-info":[{"award-number":["22361132536"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22404066"],"award-info":[{"award-number":["22404066"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22476067"],"award-info":[{"award-number":["22476067"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The health risks posed by chloroneb (CHL), a representative fungicide, in agricultural products and the environment necessitate rapid detection methods. Here, computational chemistry-aided rational hapten design yielded a high-performance monoclonal antibody against CHL (IC50 = 2.63 ng\/mL), guided by analysis of its spatial conformation and electronic properties. A dual-T-line gold nanoparticle-based immunochromatographic assay (AuNP-ICA) was established, achieving 2.6-fold higher sensitivity and a 2.9-fold higher upper detection limit compared with traditional single-T-line formats. The assay enables rapid (&amp;lt;20 min) on-site detection of CHL in lake water, soil, tea, soybeans, and mangoes, with limits of detection ranging from 2.51 to 4.54 \u03bcg\/kg. Recovery and blind sample analyses demonstrated excellent agreement with instrumental methods. This work not only provides a practical tool for field detection of CHL but also establishes a generalizable technological framework that integrates rational hapten design with a dual-T-line configuration for developing sensitive immunoassays targeting other small-molecule contaminants.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c06923","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T21:02:37Z","timestamp":1788469357000},"page":"28298-28313","source":"Crossref","is-referenced-by-count":0,"title":["A Dual-T-Line Immunochromatographic\nAssay Based on\nComputational Chemistry-Guided Hapten Design for Rapid On-Site Detection\nof Chloroneb in Agricultural and Environmental Samples"],"prefix":"10.1021","volume":"74","author":[{"given":"Feng","family":"He","sequence":"first","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, P. R. China, 214122"]},{"name":"Jiangnan University , , ,","place":["Wuxi, People\u2019s Republic of China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1930-7814","authenticated-orcid":true,"given":"Lingling","family":"Guo","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, P. R. China, 214122"]},{"name":"Jiangnan University , , ,","place":["Wuxi, People\u2019s Republic of China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5332-7101","authenticated-orcid":true,"given":"Maozhong","family":"Sun","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, P. R. China, 214122"]},{"name":"Jiangnan University , , ,","place":["Wuxi, People\u2019s Republic of China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liqiang","family":"Liu","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, P. R. China, 214122"]},{"name":"Jiangnan University , , ,","place":["Wuxi, People\u2019s Republic of China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9453-7703","authenticated-orcid":true,"given":"Liguang","family":"Xu","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, P. R. China, 214122"]},{"name":"Jiangnan University , , ,","place":["Wuxi, People\u2019s Republic of China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2724-8722","authenticated-orcid":true,"given":"Hua","family":"Kuang","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, P. R. China, 214122"]},{"name":"Jiangnan University , , ,","place":["Wuxi, People\u2019s Republic of China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5639-7102","authenticated-orcid":true,"given":"Chuanlai","family":"Xu","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, P. R. China, 214122"]},{"name":"Jiangnan University , , ,","place":["Wuxi, People\u2019s Republic of China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-6133-3828","authenticated-orcid":true,"given":"Xinxin","family":"Xu","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, P. R. China, 214122"]},{"name":"Jiangnan University , , ,","place":["Wuxi, People\u2019s Republic of China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,3]]},"reference":[{"issue":"4","key":"2026091609100422800_cit1","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1007\/s00604-021-04759-4","article-title":"Cobalt(II)-ion-exchanged Zn-bio-MOF-1\nderived CoS\/ZnS composites modified electrochemical sensor for chloroneb\ndetection by differential pulse voltammetry","volume":"188","author":"Duan","year":"2021","journal-title":"Microchim. Acta"},{"key":"2026091609100422800_cit2","doi-asserted-by":"publisher","first-page":"2952","DOI":"10.1021\/es0486418","article-title":"Gas-Phase\nConcentrations of Current-Use Pesticides\nin Iowa","volume":"39","author":"Peck","year":"2005","journal-title":"Environ. Sci. Technol."},{"key":"2026091609100422800_cit3","doi-asserted-by":"publisher","first-page":"139597","DOI":"10.1016\/j.chemosphere.2023.139597","article-title":"Polybrominated\ndiphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), organochlorine\npesticides (OCPs) in human breast milk from Colombia: A probabilistic\nrisk assessment approach","volume":"339","author":"Torres-Moreno","year":"2023","journal-title":"Chemosphere"},{"key":"2026091609100422800_cit4","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1007\/s10337-018-3483-7","article-title":"Method\nValidation Using Normal and Weighted Linear Regression Models for\nQuantification of Pesticides in Mango (Mangifera indica L.) Samples","volume":"81","author":"Martins","year":"2018","journal-title":"Chromatographia"},{"key":"2026091609100422800_cit5","doi-asserted-by":"publisher","first-page":"104617","DOI":"10.1016\/j.yrtph.2020.104617","article-title":"Comparison of the potential\nmechanisms for hepatotoxicity of p-dialkoxy chlorobenzenes in rat\nprimary hepatocytes for read-across","volume":"113","author":"Nakagawa","year":"2020","journal-title":"Regul.\nToxicol. Pharmacol."},{"key":"2026091609100422800_cit6","unstructured":"European Commission.\n          \n          EU pesticides database. https:\/\/ec.europa.eu\/food\/plant\/pesticides\/eu-pesticides-database\/start\/screen\/active-substances\/details\/704, 2009. (accessed 23 September 2025)."},{"key":"2026091609100422800_cit7","doi-asserted-by":"publisher","first-page":"5896","DOI":"10.1016\/j.chroma.2010.07.065","article-title":"Solvent-based de-emulsification\ndispersive liquid-liquid microextraction combined with gas chromatography-mass\nspectrometry for determination of trace organochlorine pesticides\nin environmental water samples","volume":"1217","author":"Zacharis","year":"2010","journal-title":"J. Chromatogr.\nA"},{"issue":"1","key":"2026091609100422800_cit8","doi-asserted-by":"publisher","first-page":"1991","DOI":"10.12681\/jhvms.22941","article-title":"Health\nRisks associated with residual pesticide levels in fish reared in\npurified wastewater from slaughterhouse","volume":"71","author":"Pelic\u0301","year":"2020","journal-title":"J.\nHellenic Vet. Med. Soc."},{"key":"2026091609100422800_cit9","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1016\/j.talanta.2016.08.015","article-title":"An effective method for pesticide residues determination\nin tobacco by GC-MS\/MS and UHPLC-MS\/MS employing acetonitrile extraction\nwith low-temperature precipitation and d-SPE clean-up","volume":"161","author":"Bernardi","year":"2016","journal-title":"Talanta"},{"key":"2026091609100422800_cit10","doi-asserted-by":"publisher","first-page":"14967","DOI":"10.1021\/acs.jafc.3c03263","article-title":"Generating Monoclonal Antibodies against Buprofezin\nand Developing\nImmunoassays for Its Residue Detection in Tea Samples","volume":"71","author":"Xu","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"2026091609100422800_cit11","doi-asserted-by":"publisher","first-page":"27669","DOI":"10.1021\/acs.jafc.5c05567","article-title":"Rational Hapten Design\nfor the Immunochromatographic\nAssay of Yohimbine, an Emerging Adulterant in Food","volume":"73","author":"Chen","year":"2025","journal-title":"J. Agric. Food Chem."},{"key":"2026091609100422800_cit12","doi-asserted-by":"publisher","first-page":"15441","DOI":"10.1021\/acs.analchem.5c02580","article-title":"Hapten Design\nand Antibody\nProduction for a Highly Sensitive Broad-Spectrum Immunoassay for Detection\nof Seven Organochlorine Pollutants in Soil and Oilfield Chemicals","volume":"97","author":"Li","year":"2025","journal-title":"Anal. Chem."},{"key":"2026091609100422800_cit13","doi-asserted-by":"publisher","first-page":"19869","DOI":"10.1021\/acs.analchem.4c02962","article-title":"Antibody Recognition\nProfile-Aided Hapten Design to Modulate Antibody Generation with Anticipated\nPerformance for Immunoassay Development","volume":"96","author":"Li","year":"2024","journal-title":"Anal.\nChem."},{"key":"2026091609100422800_cit14","doi-asserted-by":"publisher","first-page":"13978","DOI":"10.1021\/acs.jafc.4c02855","article-title":"Epitope-Shared Hapten Enhancing Antibody\nPolyreactivity for Simultaneous\nImmunochromatography of Oxyphenisatin Adulterants","volume":"72","author":"Liu","year":"2024","journal-title":"J. Agric. Food Chem."},{"key":"2026091609100422800_cit15","doi-asserted-by":"publisher","first-page":"27059","DOI":"10.1021\/acs.jafc.5c07301","article-title":"Development of an\nUltrasensitive\nMonoclonal Antibody-Based Time-Resolved Fluorescence Immunochromatographic\nAssay for Mebendazole Detection","volume":"73","author":"Chen","year":"2025","journal-title":"J. Agric. Food\nChem."},{"key":"2026091609100422800_cit16","doi-asserted-by":"publisher","first-page":"140080","DOI":"10.1016\/j.jhazmat.2025.140080","article-title":"Multiplex immunochromatographic assay for simultaneous detection\nof triclosan, triclocarban and chlorophene in environmental samples","volume":"499","author":"Li","year":"2025","journal-title":"J. Hazard. Mater."},{"key":"2026091609100422800_cit17","doi-asserted-by":"publisher","first-page":"133221","DOI":"10.1016\/j.jhazmat.2023.133221","article-title":"Monospecific\nand ultrasensitive detection of ofloxacin: A computational chemistry-assisted\nhapten screening strategy and analysis of molecular recognition mechanism","volume":"465","author":"Xiao","year":"2024","journal-title":"J. Hazard. Mater."},{"key":"2026091609100422800_cit18","doi-asserted-by":"publisher","first-page":"129931","DOI":"10.1016\/j.jhazmat.2022.129931","article-title":"Broad-specificity antibody profiled\nby hapten prediction and its application in immunoassay for fipronil\nand major metabolites","volume":"441","author":"Li","year":"2023","journal-title":"J. Hazard. Mater."},{"key":"2026091609100422800_cit19","doi-asserted-by":"publisher","first-page":"17859","DOI":"10.1021\/acs.analchem.4c04537","article-title":"Hapten Synthesis,\nAntibody Generation, and Immunoassay Development for Linezolid Therapeutic\nMonitoring","volume":"96","author":"Burkin","year":"2024","journal-title":"Anal. Chem."},{"key":"2026091609100422800_cit20","doi-asserted-by":"publisher","first-page":"140626","DOI":"10.1016\/j.jhazmat.2025.140626","article-title":"Development of a broad-specificity\nand dual T-line architecture immunochromatographic\nassay based on computational chemistry-assisted hapten screening for\nthe detection of mesotrione, its metabolites and sulcotrione","volume":"502","author":"He","year":"2026","journal-title":"J. Hazard. Mater."},{"key":"2026091609100422800_cit21","doi-asserted-by":"publisher","first-page":"134067","DOI":"10.1016\/j.jhazmat.2024.134067","article-title":"Generation\nof a highly specific recombinant full-length antibody\nfor detecting ethirimol in fruit and environmental water","volume":"469","author":"Lei","year":"2024","journal-title":"J. Hazard. Mater."},{"key":"2026091609100422800_cit22","doi-asserted-by":"publisher","first-page":"138993","DOI":"10.1016\/j.jhazmat.2025.138993","article-title":"Three-color\ntriple platform for point-of-care detection of three antifungal azoles\nin livestock wastewater and sludge","volume":"495","author":"Li","year":"2025","journal-title":"J. Hazard.\nMater."},{"key":"2026091609100422800_cit23","doi-asserted-by":"publisher","first-page":"126533","DOI":"10.1016\/j.jhazmat.2021.126533","article-title":"An immunochromatographic\nsensor for ultrasensitive and direct detection of histamine in fish","volume":"419","author":"Zeng","year":"2021","journal-title":"J. Hazard. Mater."},{"key":"2026091609100422800_cit24","doi-asserted-by":"publisher","first-page":"2739","DOI":"10.1021\/cr2001178","article-title":"Gold nanoparticles\nin chemical and biological sensing","volume":"112","author":"Saha","year":"2012","journal-title":"Chem. Rev."},{"key":"2026091609100422800_cit25","doi-asserted-by":"publisher","first-page":"3788","DOI":"10.1038\/s41596-020-0357-x","article-title":"Tutorial: design and fabrication of nanoparticle-based lateral-flow\nimmunoassays","volume":"15","author":"Parolo","year":"2020","journal-title":"Nat. Protoc."},{"key":"2026091609100422800_cit26","doi-asserted-by":"publisher","first-page":"17843","DOI":"10.1021\/acs.analchem.2c03592","article-title":"Minimum Distance Between\nTwo Epitopes in Sandwich Immunoassays for Small Molecules","volume":"94","author":"Bai","year":"2022","journal-title":"Anal. Chem."},{"key":"2026091609100422800_cit27","doi-asserted-by":"publisher","first-page":"138912","DOI":"10.1016\/j.foodchem.2024.138912","article-title":"Visual authenticating hazardous adulterant phenolphthalein\nin slimming foods: Target-mimicking hapten epitope improved immunoassay","volume":"446","author":"Liu","year":"2024","journal-title":"Food Chem."},{"key":"2026091609100422800_cit28","doi-asserted-by":"publisher","first-page":"29119","DOI":"10.1021\/acs.jafc.5c09933","article-title":"Highly Specific Monoclonal Antibodies\nCombined with A Multiplex Quantum Dot Microsphere-based Fluorescence\nLateral Flow Immunochromatographic Assay for Simultaneous Detection\nof Vanillin, Methyl Vanillin, and Ethyl Vanillin in Food Samples","volume":"73","author":"He","year":"2025","journal-title":"J. Agric. Food Chem."},{"key":"2026091609100422800_cit29","doi-asserted-by":"publisher","first-page":"137168","DOI":"10.1016\/j.jhazmat.2025.137168","article-title":"Hapten prediction, monoclonal antibody preparation,\nand development\nof an immunochromatographic assay for the detection of fenamiphos","volume":"487","author":"Lu","year":"2025","journal-title":"J. Hazard. Mater."},{"key":"2026091609100422800_cit30","doi-asserted-by":"publisher","first-page":"139552","DOI":"10.1016\/j.jhazmat.2025.139552","article-title":"Pharmacophore-guided\nproduction of high-affinity antibodies for rapid\ndetection of antipyrine in food and environment","volume":"497","author":"Pan","year":"2025","journal-title":"J. Hazard. Mater."},{"key":"2026091609100422800_cit31","doi-asserted-by":"publisher","first-page":"3160","DOI":"10.1021\/acs.jafc.3c07766","article-title":"Antibody Production,\nImmunoassay Establishment, and Specificity Study for Flunixin and\n5-Hydroxyflunixin","volume":"72","author":"Li","year":"2024","journal-title":"J. Agric. Food Chem."},{"key":"2026091609100422800_cit32","doi-asserted-by":"publisher","first-page":"134483","DOI":"10.1016\/j.foodchem.2022.134483","article-title":"Development\nof a GNP-based lateral flow immunoassay for the detection of isoprothiolane\nin rice samples","volume":"404","author":"Liu","year":"2023","journal-title":"Food Chem."},{"key":"2026091609100422800_cit33","doi-asserted-by":"publisher","first-page":"11170","DOI":"10.1021\/acs.jafc.8b03834","article-title":"Generic\nHapten Synthesis, Broad-Specificity Monoclonal Antibodies Preparation,\nand Ultrasensitive ELISA for Five Antibacterial Synergists in Chicken\nand Milk","volume":"66","author":"Li","year":"2018","journal-title":"J. Agric. Food Chem."},{"key":"2026091609100422800_cit34","doi-asserted-by":"publisher","first-page":"340699","DOI":"10.1016\/j.aca.2022.340699","article-title":"The influence of hapten spacer arm\nlength on antibody response and immunoassay development","volume":"1239","author":"Bai","year":"2023","journal-title":"Anal. Chim. Acta"},{"key":"2026091609100422800_cit35","doi-asserted-by":"publisher","first-page":"144723","DOI":"10.1016\/j.foodchem.2025.144723","article-title":"Computational\nchemistry-based hapten design and antibody production\nfor the immunochromatographic assay of maleic hydrazide in food and\nenvironmental samples","volume":"487","author":"He","year":"2025","journal-title":"Food Chem."},{"key":"2026091609100422800_cit36","doi-asserted-by":"publisher","first-page":"137634","DOI":"10.1016\/j.jhazmat.2025.137634","article-title":"Visual immunosensor\nassay with double T line for on-site\nsensitive fipronil pesticide detection in water, soil and honeysuckle","volume":"489","author":"Kou","year":"2025","journal-title":"J. Hazard. Mater."},{"key":"2026091609100422800_cit37","doi-asserted-by":"publisher","first-page":"134100","DOI":"10.1016\/j.jhazmat.2024.134100","article-title":"Quantitative immunochromatographic\nassay for rapid and cost-effective on-site detection of benzo[a]pyrene\nin oilfield chemicals","volume":"469","author":"Li","year":"2024","journal-title":"J. Hazard. Mater"},{"key":"2026091609100422800_cit38","doi-asserted-by":"publisher","first-page":"139999","DOI":"10.1016\/j.foodchem.2024.139999","article-title":"General hapten skeleton\nmotivated duplex-immunoassay for emergent bisoxatin adulterants in\nslimming foods","volume":"456","author":"Liu","year":"2024","journal-title":"Food Chem."},{"key":"2026091609100422800_cit39","doi-asserted-by":"publisher","first-page":"4819","DOI":"10.1021\/acs.jafc.7b00243","article-title":"Matrix\nEffects and Interferences of Different Citrus Fruit Coextractives\nin Pesticide Residue Analysis Using Ultrahigh-Performance Liquid Chromatography-High-Resolution\nMass Spectrometry","volume":"65","author":"Besil","year":"2017","journal-title":"J. Agric. Food Chem."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c06923","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/36\/28298\/68293155\/acs.jafc.6c06923.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/36\/28298\/68293155\/acs.jafc.6c06923.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T13:25:04Z","timestamp":1789565104000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/36\/28298\/5413963\/A-Dual-T-Line-Immunochromatographic-Assay-Based-on"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,3]]},"references-count":39,"journal-issue":{"issue":"36","published-online":{"date-parts":[[2026,9,3]]},"published-print":{"date-parts":[[2026,9,16]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c06923","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,3]]}}}