Development of a Simple Pretreatment Immunoassay Based on an Organic Solvent-Tolerant Nanobody for the Detection of Carbofuran in Vegetable and Fruit Samples
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials and Reagents
2.2. Construction of Phage Display Nanobody Library
2.3. Selection and Identification of Anti-Carbofuran Phage Clones
2.4. Expression and Purification of Nanobody Protein
2.5. Stability Analysis of Anti-Carbofuran Nanobody
2.6. Nanobody-Based Indirect Competitive ELISA
2.7. Immunoassay Validation
3. Results and Discussion
3.1. Library Construction and Selection of Anti-Carbofuran Phage Clones
3.2. Preparation and Stability Analysis of Anti-Carbofuran Nanobody
3.3. Nanobody-Based Ic-ELISA
3.4. Sample Analysis by ic-ELISA and UPLC–MS/MS
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Analogues | Molecular Structural | IC50 (ng/mL) | Cross-Reactivity (%) |
---|---|---|---|
Carbofuran | 7.27 | 100 | |
Benfuracarb | 142.51 | 5.1 | |
Fenobucarb | 204.6 | 3.5 | |
Carbosulfan | 280.93 | 2.6 | |
3-Hydroxycarbofuran | 366.05 | 2.0 | |
Isoprocarb | 1351.82 | 0.5 | |
Carbaryl | >2000 | <0.1 | |
Aldicarb | >2000 | <0.1 | |
Methomyl | >2000 | <0.1 | |
Pirimicarb | >2000 | <0.1 | |
Mercaptodimethur | >2000 | <0.1 | |
Tsumacide | >2000 | <0.1 |
Added (mg/kg) | Chinese Cabbage | Cucumber | Orange | ||||||
---|---|---|---|---|---|---|---|---|---|
Found ± SD (mg/kg) | Recovery (%) | CV (%) | Found ± SD (mg/kg) | Recovery (%) | CV (%) | Found ± SD (mg/kg) | Recovery (%) | CV (%) | |
10 | 10.27 ± 0.34 | 102.7 | 3.33 | 9.73 ± 0.24 | 97.35 | 2.52 | 8.47 ± 0.2 | 84.74 | 2.35 |
20 | 18.44 ± 0.54 | 92.25 | 2.61 | 20.45 ± 0.31 | 102.26 | 1.50 | 16.46 ± 0.22 | 82.31 | 1.21 |
50 | 51.96 ± 0.75 | 103.92 | 1.44 | 46.24 ± 0.51 | 92.48 | 1.11 | 43.77 ± 0.91 | 87.54 | 20.76 |
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Zhang, J.-r.; Wang, Y.; Dong, J.-x.; Yang, J.-y.; Zhang, Y.-q.; Wang, F.; Si, R.; Xu, Z.-l.; Wang, H.; Xiao, Z.-l.; et al. Development of a Simple Pretreatment Immunoassay Based on an Organic Solvent-Tolerant Nanobody for the Detection of Carbofuran in Vegetable and Fruit Samples. Biomolecules 2019, 9, 576. https://doi.org/10.3390/biom9100576
Zhang J-r, Wang Y, Dong J-x, Yang J-y, Zhang Y-q, Wang F, Si R, Xu Z-l, Wang H, Xiao Z-l, et al. Development of a Simple Pretreatment Immunoassay Based on an Organic Solvent-Tolerant Nanobody for the Detection of Carbofuran in Vegetable and Fruit Samples. Biomolecules. 2019; 9(10):576. https://doi.org/10.3390/biom9100576
Chicago/Turabian StyleZhang, Jin-ru, Yu Wang, Jie-xian Dong, Jin-yi Yang, Yu-qi Zhang, Feng Wang, Rui Si, Zhen-lin Xu, Hong Wang, Zhi-li Xiao, and et al. 2019. "Development of a Simple Pretreatment Immunoassay Based on an Organic Solvent-Tolerant Nanobody for the Detection of Carbofuran in Vegetable and Fruit Samples" Biomolecules 9, no. 10: 576. https://doi.org/10.3390/biom9100576