Development of indirect competitive ELISA and CLEIA for quantitative analysis of melatonin in health products

Wu, Longjiang; Isah Bindawa, Murtala; Zhang, Siran; Dang, Mei; Zhang, Xiaoying

doi:10.1039/d3ay01182h

Anal. Methods

2023

DOI: 10.1039/d3ay01182h

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Development of indirect competitive ELISA and CLEIA for quantitative analysis of melatonin in health products

Longjiang Wu,

Murtala Isah Bindawa,

Siran Zhang

et al.

Abstract: Melatonin (MT), as a hormone regulating the rhythm of sleep, is widely used in health products. However, illicit and excessive use of MT might cause undesirable effects. Therefore, it is...

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Cited by 6 publications

(2 citation statements)

References 29 publications

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“…Chemical structure and detection methods of AAI: (A) Structure representation of AAI within the Aristolochia genus. (B) The detection ranges of previously developed methods for detecting AAI. − Competition inhibition curve (I) and standard curve (II) of ic-ELISA (C) and dc-QD-FLISA (D) for AAI detection.…”

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confidence: 99%

“…The development of the ic-ELISA and dc-QD-FLISA followed established protocols briefed in the Supporting Information. ,, Subsequently, spike recovery experiments were conducted. In the context of the developed ic-ELISA, it was observed that the average recoveries for AAI in herbal medicines and traditional Chinese medicines consistently fell within the range from 85.7% to 111.89%.…”

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confidence: 99%

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Quantum Dot-Based Immunoassays: Unraveling Sensitivity Discrepancies and Charting Future Frontiers

Dang,

Wu,

Jin

et al. 2024

Anal. Chem.

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The 2023 Nobel Prize in Chemistry honors the groundbreaking contributions of Alexei Ekimov, Louis Brus, and Moungi Bawendi to the field of quantum dots (QDs). In this spirit, we developed a direct competitive QD fluorescence immunoassay (dc-QD-FLISA) to detect aristolochic acid type I (AAI), a potent carcinogen found in herbal remedies. Unexpectedly, the dc-QD-FLISA exhibited lower sensitivity than that of an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), contrary to our initial expectations. This discrepancy in the sensitivity prompted a comprehensive analysis of the entire experimental process. We propose that steric hindrance between QDs and antigen-binding sites on antibodies may significantly diminish the binding efficiency, reducing sensitivity within the dc-QD-FLISA method. Furthermore, issues such as buffer conditions, antibody handling, and separation methods are also contributing factors. We recommend site-directed QD modification and stringent consideration of the experimental conditions. This study not only provides insights into QD-based immunoassays but also highlights the need for future advancements in immunoassay technology in terms of augmenting sensitivity and specificity, potentially revolutionizing disease diagnosis, biomarker discovery, and biomedical research.

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confidence: 99%

mentioning

confidence: 99%

Quantum Dot-Based Immunoassays: Unraveling Sensitivity Discrepancies and Charting Future Frontiers

Dang,

Wu,

Jin

et al. 2024

Anal. Chem.

Self Cite

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DNA-directed immobilization fluorescent immunoarray for multiplexed antibiotic residue determination in milk

Guercetti,

Pascual,

Aviñó

et al. 2024

Anal Bioanal Chem

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The presence of antibiotic residues in cow’s milk entails high risk for consumers, the dairy industry, and the environment. Therefore, the development of highly specific and sensitive screening tools for the rapid and cost-effective identification of traces of these compounds is urgently needed. A multiplexed screening platform utilizing DNA-directed immobilization (DDI) was developed aiming to detect three classes of antibiotic residues (fluoroquinolones, sulfonamides, and tylosin) prevalently found in milk. Throughout this work, each oligonucleotide sequence was conjugated to a different hapten molecule, while the three complementary strands were immobilized in 24 independent microarray chips on a single glass slide. First, the array was incubated with the pool of hapten-oligonucleotide conjugate site encoded the signal through DNA hybridization. Next, commercial milk samples were incubated with the cocktail of monoclonal antibodies following a secondary fluorophore-labeled antibody which was required for fluorescent readout. Direct sample detection was achieved in milk diluting 20 times in assay buffer. The limits of detection (LODs) reached were 1.43 µg kg−1, 1.67 µg kg−1, and 0.89 µg kg−1 for TYLA, STZ, and CIP, respectively, which represented in raw milk 7.15 µg kg−1, 8.35 µg kg−1, and 4.45 µg kg−1 for TYLA, STZ, and CIP, respectively, that are below the EU regulatory limits. Cross-reactivity profiles were evaluated against the family of structurally related antibiotics in order to demonstrate the capability to detect antibiotics from the same family of compounds. A pre-validation study was performed by spiking 20 blind samples above and below the maximum residue limits established by the EU guidelines. The system was successfully implemented towards randomized sample classification as compliant or non-compliant. The proposed DDI-based immunoarray provides a fast and cost-effective alternative to obtain semi-quantitative information about the presence of three veterinary residues simultaneously in milk samples. Graphical Abstract Antibody fluorescent microarray based on DDI. The figure shows the main steps involved in the immunoassay. First, the printing of the oligo N4-6down probe over the glass slide, followed by an incubation with a complementary strand conjugated to the hapten and finally the selective recognition using monoclonal antibodies and fluorescent quantification.

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Development and evaluation of an immunoassay for the quantification of N-acetylneuraminic acid (Neu5Ac) in foods and biosamples

Wu,

Deng,

Isah

et al. 2024

Food Chemistry

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Development of indirect competitive ELISA and CLEIA for quantitative analysis of melatonin in health products

Abstract: Melatonin (MT), as a hormone regulating the rhythm of sleep, is widely used in health products. However, illicit and excessive use of MT might cause undesirable effects. Therefore, it is...

Cited by 6 publications

References 29 publications

Quantum Dot-Based Immunoassays: Unraveling Sensitivity Discrepancies and Charting Future Frontiers

Quantum Dot-Based Immunoassays: Unraveling Sensitivity Discrepancies and Charting Future Frontiers

DNA-directed immobilization fluorescent immunoarray for multiplexed antibiotic residue determination in milk

Development and evaluation of an immunoassay for the quantification of N-acetylneuraminic acid (Neu5Ac) in foods and biosamples

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