A Highly Sensitive and Group-Specific Enzyme-Linked Immunosorbent Assay for the Detection of AFB[sub.1] in Agriculture and Aquiculture Products
Aflatoxins (AFs) including AFB[sub.1], AFB[sub.2], AFG[sub.1] and AFG[sub.2] are widely found in agriculture products, and AFB[sub.1] is considered one of the most toxic and harmful mycotoxins. Herein, a highly sensitive (at the pg mL[sup.−1] level) and group-specific enzyme-linked immunosorbent ass...
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Veröffentlicht in: | Molecules (Basel, Switzerland) Switzerland), 2024-05, Vol.29 (10) |
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Zusammenfassung: | Aflatoxins (AFs) including AFB[sub.1], AFB[sub.2], AFG[sub.1] and AFG[sub.2] are widely found in agriculture products, and AFB[sub.1] is considered one of the most toxic and harmful mycotoxins. Herein, a highly sensitive (at the pg mL[sup.−1] level) and group-specific enzyme-linked immunosorbent assay (ELISA) for the detection of AFB[sub.1] in agricultural and aquiculture products was developed. The AFB[sub.1] derivative containing a carboxylic group was synthesized and covalently linked to bovine serum albumin (BSA). The AFB[sub.1]-BSA conjugate was used as an immunogen to immunize mice. A high-quality monoclonal antibody (mAb) against AFB[sub.1] was produced by hybridoma technology, and the mAb-based ELISA for AFB[sub.1] was established. IC[sub.50] and limit of detection (LOD) of the ELISA for AFB[sub.1] were 90 pg mL[sup.−1] and 18 pg mL[sup.−1], respectively. The cross-reactivities (CRs) of the assay with AFB[sub.2], AFG[sub.1], and AFG[sub.2] were 23.6%, 42.5%, and 1.9%, respectively, revealing some degree of group specificity. Corn flour, wheat flour, and crab roe samples spiked with different contents of AFB[sub.1] were subjected to ELISA procedures. The recoveries and relative standard deviation (RSD) of the ELISA for AFB[sub.1] in spiked samples were 78.3–116.6% and 1.49–13.21% (n = 3), respectively. Wheat flour samples spiked with the mixed AF (AFB[sub.1], AFB[sub.2], AFG[sub.1], AFG[sub.2]) standard solution were measured by ELISA and LC-MS/MS simultaneously. It was demonstrated that the proposed ELISA can be used as a screening method for evaluation of AFs (AFB[sub.1], AFB[sub.2], AFG[sub.1], AFG[sub.2]) in wheat flour samples. |
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ISSN: | 1420-3049 1420-3049 |
DOI: | 10.3390/molecules29102280 |