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Investigation of the Small Size of Nanobodies for a Sensitive Fluorescence Polarization Immunoassay for Small Molecules: 3-Phenoxybenzoic Acid, an Exposure Biomarker of Pyrethroid Insecticides as a Model

文献类型： 外文期刊

作者： Wang, Yulong 1 ; Li, Zhenfeng 2 ; Barnych, Bogdan 2 ; Huo, Jingqian 2 ; Wan, Debin 2 ; Vasylieva, Natalia 2 ; Xu, Junl 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Jiangsu Key Lab Food Qual & Safety, State Key Lab Cultivat Base, Minist Sci & Technol,Inst Food Safety & Nutr, Nanjing 210014, Jiangsu, Peoples R China

2.Univ Calif Davis, Dept Entomol & Nematol, Davis, CA 95616 USA

3.Univ Calif Davis, UCD Comprehens Canc Ctr, Davis, CA 95616 USA

4.Jiangsu Univ, Sch Food & Biol Engn, Zhenjiang 212013, Jiangsu, Peoples R China

5.Agr Univ Hebei, Coll Plant Protect, Baoding 071001, Peoples R China

关键词： fluorescence polarization immunoassay; nano body; insecticide; 3-phenoxybenzoic acid; fluorescein

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：5.279； 五年影响因子：5.269 ）

ISSN： 0021-8561

年卷期： 2019 年 67 卷 41 期

页码：

收录情况： SCI

摘要： Limited reports on the use of nanobodies (Nbs) in fluorescence polarization immunoassay (FPIA) aroused us to explore if the small size of Nbs is a drawback for the development of sensitive FPIA to small molecular compounds, particularly since FPIA is a technology strongly dependent on molecular weight. In the present work, three different molecular weight Nbs against 3-phenoxybenzoic acid (3-PBA), an exposure biomarker of pyrethroid insecticides, including bare Nbs (15 kDa), Nbs-Avidin (Nbs-AV, 60 kDa), and Nbs-Alkaline phosphatase (Nbs-AP, 130 kDa) were specifically generated to cover distinct regions on the polarization and molecular weight relationship curve for a fluorescein tracer. In competitive FPIA, similar halfmaximal inhibitory concentrations (IC50) of 3-PBA of 16.4, 12.2, and 14.8 ng mL(-1) were obtained for Nbs, Nbs-AV, and Nbs-AP, respectively, indicating that the size of Nbs in the range tested had no significant effect on the sensitivity of the resulting competitive FPIA. An IC50 of 20.2 ng mL(-1) for an anti-3-PBA polyconal antibody based FPIA further demonstrated the performance of Nbs, which was comparable to that of traditional antibodies in FPIA. Spike-recovery studies showed good and reproducible recovery of 3-PBA in urine samples, demonstrating the applicability of Nb-based FPIA. Overall, our results show that Nb-based FPIA achieves sensitivity levels of FPIA based on conventional antibodies and further indicate that Nb absolutely meets the sensitivity requirement of FPIA.

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