A Novel Full-length IgG Recombinant Antibody Highly Specific to Clothianidin and Its Application in Immunochromatographic Assay
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents and Materials
2.2. Hybridoma Screening
2.3. Sequencing and Analysis of Antibody VR Genes
2.4. Cloning Antibody Genes into Full-Length RAbs
2.4.1. Construction of Expression Vectors
2.4.2. Production of Full-Length RAbs
2.5. Functional Analyses of Full-Length RAbs
2.5.1. Heterologous ic-ELISAs
2.5.2. Non-Competitive SPR
2.6. Development of Full-Length RAb-Based GICA for Clothianidin Detection
2.6.1. Selectivity and Sensitivity Experiment
2.6.2. Sample Matrix Effect Analysis
2.6.3. Analysis of the Spiked Samples
3. Results and Discussion
3.1. Hybridoma Screening
3.2. Sequencing of VR Genes and Expression of Full-Length IG RAbs
3.3. Heterologous ic-ELISAs for Functional Characterization of Full-Length RAbs
3.4. Non-Competitive SPR Assay for Functional Characterization of Full-Length RAbs
3.5. Application of a Highly Specific RAb in a GICA for Clothianidin Detection
3.5.1. Specificity and Sensitivity of GICA
3.5.2. Matrix Effects
3.5.3. Application of the GICA in Spiked Samples
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Neonicotinoid Analogues | PcAb [22] | McAb [23] | McAb [35] | 5C3-McAb in This Study | 5C3-RAb in This Study | 1F7-McAb in This Study | 1F7-RAb in This Study |
---|---|---|---|---|---|---|---|
| 100 a (46) b | 100 (4.4) | 100 (25.6) | 100 (13.16) | 100 (5.22) | 100 (5.38) | 100 (4.62) |
| 11.8 c | 64 | 47.8 | 42.4 | 56 | <0.1 | <0.1 |
| 0.8 | <0.1 | <0.03 | <0.1 | <0.1 | <0.1 | <0.1 |
| <0.05 | <0.1 | <0.03 | <0.1 | <0.1 | <0.1 | <0.1 |
| <0.05 | <0.1 | <0.03 | <0.1 | <0.1 | <0.1 | <0.1 |
| <0.05 | <0.1 | <0.03 | <0.1 | <0.1 | <0.1 | <0.1 |
| <0.05 | <0.1 | <0.03 | <0.1 | <0.1 | <0.1 | <0.1 |
| <0.05 | <0.1 | <0.03 | <0.1 | <0.1 | <0.1 | <0.1 |
Sample | Spiked Concentration (μg/L or ng/g) | Dilution Time d | Results (n = 3) | |
---|---|---|---|---|
Batch 1 e | Batch 2 | |||
River water | 0 | 2 | −/−/− | −/−/− |
5 | ±/±/± | ±/±/± | ||
10 | +/+/+ | +/+/+ | ||
20 | +/+/+ | +/+/+ | ||
Soil | 0 | 4 | −/−/− | −/−/− |
10 | ±/±/± | ±/±/± | ||
20 | +/+/+ | +/+/+ | ||
40 | +/+/+ | +/+/+ | ||
Tomato | 0 | 6 | −/−/− | −/−/− |
15 | ±/±/± | ±/±/± | ||
30 | +/+/+ | +/+/+ | ||
60 | +/+/+ | +/+/+ | ||
Orange | 0 | 6 | −/−/− | −/−/− |
15 | ±/±/± | ±/±/± | ||
30 | +/+/+ | +/+/+ | ||
60 | +/+/+ | +/+/+ |
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Chang, Y.; Chen, Y.; Jiao, S.; Lu, X.; Fang, Y.; Liu, Y.; Zhao, Y.; Zhan, X.; Zhu, G.; Guo, Y. A Novel Full-length IgG Recombinant Antibody Highly Specific to Clothianidin and Its Application in Immunochromatographic Assay. Biosensors 2022, 12, 233. https://doi.org/10.3390/bios12040233
Chang Y, Chen Y, Jiao S, Lu X, Fang Y, Liu Y, Zhao Y, Zhan X, Zhu G, Guo Y. A Novel Full-length IgG Recombinant Antibody Highly Specific to Clothianidin and Its Application in Immunochromatographic Assay. Biosensors. 2022; 12(4):233. https://doi.org/10.3390/bios12040233
Chicago/Turabian StyleChang, Yunyun, Yang Chen, Shasha Jiao, Xinying Lu, Yihua Fang, Yihua Liu, Ying Zhao, Xiuping Zhan, Guonian Zhu, and Yirong Guo. 2022. "A Novel Full-length IgG Recombinant Antibody Highly Specific to Clothianidin and Its Application in Immunochromatographic Assay" Biosensors 12, no. 4: 233. https://doi.org/10.3390/bios12040233