Development of a Biotinylated Nanobody-Based Gold Nanoparticle Immunochromatographic Assay for the Detection of Procymidone in Crops
文献类型： 外文期刊
作者： Liu, Min-Ling 1 ; He, Xiao-Ting 1 ; Xu, Zhen-Lin 1 ; Deng, Hao 2 ; Shen, Yu-Dong 1 ; Luo, Lin 1 ; Shen, Xing 1 ; Chen, Zi-Jian 5 ; Hammock, Bruce 3 ; Wang, Hong 1 ;
作者机构： 1.South China Agr Univ, Guangdong Prov Key Lab Food Qual & Safety, Guangzhou 510642, Peoples R China
2.Hainan Acad Agr Sci, Key Lab Trop Fruit & Vegetable Cold Chain Hainan, Inst Agroprod Proc & Design, Haikou 570100, Hainan, Peoples R China
3.Univ Calif Davis, Dept Entomol, Davis, CA 95616 USA
4.Univ Calif Davis, UCD Comprehens Canc Ctr, Davis, CA 95616 USA
5.Zhaoqing Univ, Sch Food & Pharmaceut Engn, Lab Qual & Safety Risk Assessment Agroprod Zhaoqi, Minist Agr & Rural Affairs, Zhaoqing 526061, Peoples R China
关键词： procymidone; nanobodies; biotinylation; streptavidin; immunochromatographic assay
期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：6.1； 五年影响因子：6.3 ）
ISSN： 0021-8561
年卷期： 2023 年 71 卷 35 期
页码：
收录情况： SCI
摘要： A heavy-chain antibody (VHH) library against procymidone (PRM) was constructed via immunizing Bactrian camels. Through careful biopanning, seven nanobodies (Nbs) with different sequences were obtained. The variability in their performance was primarily attributed to the amino acid differences in complementarity-determining region 3 (CDR3), as analyzed by molecular docking. The Nb exhibiting the highest sensitivity, named NbFM5, was biotinylated and conjugated to streptavidin-labeled gold nanoparticles to preserve the epitope's activity and prevent a decrease in sensitivity due to traditional random electrostatic adsorption. Subsequently, a simple and sensitive immunochromatographic assay (ICA) was developed for rapid detection of PRM based on biotinylated Nb (btNb). The developed btNb-ICA showed a cut-off value of 200 ng/mL for visual judgment and a half-inhibitory concentration (IC50) of 6.04 ng/mL for quantitative detection. The limit of detection (LOD) was as low as 0.88 ng/mL. The recoveries in actual samples of crops ranged from 82.2 to 117.3%, aligning well with the results obtained from GC-MS/MS (R-2 = 0.995). In summary, the developed btNb-ICA demonstrated high specificity and good accuracy for the rapid detection of PRM residues in vegetables. The total analysis time from preparing the sample to obtaining the result was less than 25 min.
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