{"title":"Development of a sensitive monoclonal antibody-based immunochromatographic strip for neomycin detection in milk","authors":"Kemeng Qin, Mingyue Ding, Chensi Zhang, Xijie Zhang, Yexuan Mao, Meng Dang, Zizhe Li, Youyi Wang, Shaohui Zhang, Yuhang Sun, Zhibin Zhang, Gaiming Zhao, Yu Li, Qian Li, Xiya Zhang","doi":"10.1080/09540105.2022.2070606","DOIUrl":null,"url":null,"abstract":"ABSTRACT To improve the detection sensitivity of neomycin (NEO) in milk, we produced a sensitive monoclonal antibody (mAb) against NEO and developed a lateral flow immunoassay based on amorphous carbon nanoparticles (ACNPs-LFA). First, we conjugated NEO to carrier protein to prepare mAbs. We obtained six mAbs: mAb 1C6, mAb 1D3, mAb 2D3, mAb 4D5, mAb 5D1, and mAb 5H1. We characterised the mAbs by indirect competitive enzyme-linked immunosorbent assay and selected the most sensitive mAb based on the half-maximal inhibitory concentration (IC50). We selected mAb 4D5 (IC50 = 0.15 ng/mL) for the development of LFA. MAb 4D5 was labelled with ACNPs (ACNPs-mAb 4D5) by electrostatic absorption. Under optimised conditions, 5.4 μg mAb 4D5 coupling with 1 mL ACNPs, NEO-OVA at concentration of 80 μg/mL, 3 μL ACNPs-mAb 4D5 were used to develop LFA. The cut-off value was 8 ng/mL. Therefore, our developed ACNPs-LFA is suitable for on-site detection of NEO residues.","PeriodicalId":12300,"journal":{"name":"Food and Agricultural Immunology","volume":null,"pages":null},"PeriodicalIF":1.7000,"publicationDate":"2022-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"4","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Food and Agricultural Immunology","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1080/09540105.2022.2070606","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CHEMISTRY, APPLIED","Score":null,"Total":0}
引用次数: 4
Abstract
ABSTRACT To improve the detection sensitivity of neomycin (NEO) in milk, we produced a sensitive monoclonal antibody (mAb) against NEO and developed a lateral flow immunoassay based on amorphous carbon nanoparticles (ACNPs-LFA). First, we conjugated NEO to carrier protein to prepare mAbs. We obtained six mAbs: mAb 1C6, mAb 1D3, mAb 2D3, mAb 4D5, mAb 5D1, and mAb 5H1. We characterised the mAbs by indirect competitive enzyme-linked immunosorbent assay and selected the most sensitive mAb based on the half-maximal inhibitory concentration (IC50). We selected mAb 4D5 (IC50 = 0.15 ng/mL) for the development of LFA. MAb 4D5 was labelled with ACNPs (ACNPs-mAb 4D5) by electrostatic absorption. Under optimised conditions, 5.4 μg mAb 4D5 coupling with 1 mL ACNPs, NEO-OVA at concentration of 80 μg/mL, 3 μL ACNPs-mAb 4D5 were used to develop LFA. The cut-off value was 8 ng/mL. Therefore, our developed ACNPs-LFA is suitable for on-site detection of NEO residues.
期刊介绍:
Food and Agricultural Immunology is an international open access journal publishing original immunological research with applications in food, agricultural, environmental and veterinary science. Submissions describing the use of immunological techniques and methods are particularly welcomed.
The journal aims to expand our understanding of the interactions at the interface of food and immune systems including studies on:
-Development of diagnostic systems – all types of ligand-based assays, e.g. antibody, aptamer
-Application of ligand-based assays for the detection or identification of molecules of interest in food science, agricultural research, veterinary investigations and clinical systems relating to food allergy or sensitivity to agricultural chemicals
-Effects of food on the immune system
-Studies on allergy and allergic reactions
-Investigations into food allergies
-Development of allergen-free food systems
-Development of novel assay formats
-Applications of assay systems to the monitoring of food items in relation to safety and labelling
-Food quality issues, e.g. speciation, adulteration and contamination
-Comparisons between different analytical techniques
The journal publishes research and review articles and is essential reading for food scientists, immunologists and all those concerned with the interaction between food and immune systems.