Development of a Highly Sensitive and Specific ic-ELISA and Lateral Flow Immunoassay for Diacetoxyscirpenol
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents and Materials
2.2. Synthesis of Hapten and Antigen
2.3. Production of mAb
2.4. Development and Optimization of ic-ELISA
2.5. Matrix Effect and Recovery
2.6. Preparation of mAb-Labeled Colloidal Gold Nanoparticles (AuNPs)
2.7. Preparation of Lateral Flow Immunoassay Strip
2.8. Extraction of Antibody Variable Region Sequences, Homology Modeling and Molecular Docking
3. Results and Discussion
3.1. Preparation and Identification of Hapten and Antigen
3.2. Production of mAb to DAS
3.3. Optimization and Development of ic-ELISA
3.4. Matrix Effect and Recovery of ic-ELISA
3.5. Development of LFIA
3.6. Recognition Mechanism of mAb to DAS
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Antibody Type | IC50 (μg/L) | Immunoassays | Reference |
---|---|---|---|
pAb | 15 | RIA | [29] |
pAb | Not mentioned | ic-ELISA | [30] |
pAb | 10 | ic-ELISA | [31] |
pAb | Not mentioned | ic-ELISA | [32] |
pAb | Not mentioned | ic-ELISA | [33] |
mAb | Not mentioned | RIA | [34] |
mAb | 5.97 | ic-ELISA | [35] |
mAb | 3.08 | competitive-type pressure-dependent immunosensor | [36] |
mAb | 0.31 | ic-ELISA | This study |
Analyte | IC50 (μg/L) | Cross-Reactivity (%) |
---|---|---|
DAS | 0.31 | 100 |
T-2 toxin | >5000 | <0.1 |
HT-2 toxin | >5000 | <0.1 |
Aflatoxin B1 | >5000 | <0.1 |
Fumonisin B1 | >5000 | <0.1 |
Deoxynivalenol | >5000 | <0.1 |
Citreoviridin | >5000 | <0.1 |
Ochratoxin A | >5000 | <0.1 |
Zearalenone | >5000 | <0.1 |
Spiked Level (μg/kg) | Mean Recovery (%) | Inter-Assay CV (%) | Intra-Assay CV (%) |
---|---|---|---|
50 | 114 | 10 | 4 |
106 | 7 | ||
109 | 13 | ||
100 | 95 | 12 | 14 |
120 | 12 | ||
124 | 4 | ||
200 | 96 | 12 | 5 |
106 | 13 | ||
104 | 7 |
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Shao, S.; Shang, W.; Bai, Y.; Dou, L.; Zhang, S.; Shen, J.; Wang, Z.; Wen, K. Development of a Highly Sensitive and Specific ic-ELISA and Lateral Flow Immunoassay for Diacetoxyscirpenol. Foods 2022, 11, 1548. https://doi.org/10.3390/foods11111548
Shao S, Shang W, Bai Y, Dou L, Zhang S, Shen J, Wang Z, Wen K. Development of a Highly Sensitive and Specific ic-ELISA and Lateral Flow Immunoassay for Diacetoxyscirpenol. Foods. 2022; 11(11):1548. https://doi.org/10.3390/foods11111548
Chicago/Turabian StyleShao, Shibei, Wenhua Shang, Yuchen Bai, Leina Dou, Suxia Zhang, Jianzhong Shen, Zhanhui Wang, and Kai Wen. 2022. "Development of a Highly Sensitive and Specific ic-ELISA and Lateral Flow Immunoassay for Diacetoxyscirpenol" Foods 11, no. 11: 1548. https://doi.org/10.3390/foods11111548