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Here, we applied a computer-aided hapten design strategy integrating density functional theory, molecular docking, and molecular dynamics simulations to engineer four AFM1 haptens with tailored spacer arms (C2, C4, C6, and phenyl). A high-affinity monoclonal antibody (10C1, IC50 = 0.015 ng\/mL) was generated. Based on this antibody, an icELISA (LOD = 0.0017 ng\/mL, meeting EU standards of 0.05 ng\/mL) and a gold nanoparticle-based LFIA (LOD = 0.3 ng\/mL, meeting China\/US\/Russia standards of 0.5 ng\/mL) were developed for laboratory and on-site detection, respectively. This strategy facilitates rational hapten optimization and efficient generation of high-affinity antibodies for AFM1 monitoring.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c07668","type":"journal-article","created":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T09:01:46Z","timestamp":1786093306000},"page":"25660-25673","source":"Crossref","is-referenced-by-count":0,"title":["Development of\nHigh-Affinity Monoclonal Antibodies\nfor Aflatoxin M1 Detection in Milk"],"prefix":"10.1021","volume":"74","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-9187-2638","authenticated-orcid":true,"given":"Zhi","family":"Zhao","sequence":"first","affiliation":[{"name":"Guangxi University , , , ,","place":["Nanning, Guangxi, China, 530004"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoge","family":"Duan","sequence":"additional","affiliation":[{"name":"Guangxi University , , , ,","place":["Nanning, Guangxi, China, 530004"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingting","family":"Yue","sequence":"additional","affiliation":[{"name":"Guangxi University , , , ,","place":["Nanning, Guangxi, China, 530004"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zeming","family":"Bu","sequence":"additional","affiliation":[{"name":"Guangxi Veterinary Drug Supervision Institute , , ,","place":["Nanning, Guangxi, China, 530001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lilan","family":"Lv","sequence":"additional","affiliation":[{"name":"Guangxi Subtropical Crops Research Institute, Ministry of Agriculture and Rural Affairs, Laboratory for Quality and Safety Risk Assessment of Agricultural Products (Nanning), Key Laboratory of Subtropical Fruits and Vegetables Quality and Safety Control , ,","place":["Nanning, China, 530001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weijing","family":"Huang","sequence":"additional","affiliation":[{"name":"Guangxi Zhuang Autonomous Region Analysis Testing Research Center , , ,","place":["Nanning, Guangxi, China, 530001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meiling","family":"Yu","sequence":"additional","affiliation":[{"name":"Guangxi University , , , ,","place":["Nanning, Guangxi, China, 530004"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiakang","family":"He","sequence":"additional","affiliation":[{"name":"Guangxi University , , , ,","place":["Nanning, Guangxi, China, 530004"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3413-3576","authenticated-orcid":true,"given":"Liang","family":"Luo","sequence":"additional","affiliation":[{"name":"Guangxi University , , , ,","place":["Nanning, Guangxi, China, 530004"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1733-5267","authenticated-orcid":true,"given":"Hailan","family":"Chen","sequence":"additional","affiliation":[{"name":"Guangxi University , , , ,","place":["Nanning, Guangxi, China, 530004"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,7]]},"reference":[{"issue":"9","key":"2026081910174841100_cit1","doi-asserted-by":"publisher","first-page":"2281","DOI":"10.1080\/10408398.2021.1960794","article-title":"Hepatotoxicity of food-borne mycotoxins:\nmolecular mechanism, anti-hepatotoxic medicines and target prediction","volume":"62","author":"Ruan","year":"2022","journal-title":"Crit. 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