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Lateral flow immunoassay for 5-hydroxyflunixin based on near-infrared fluorescence molecule as an alternative label to gold nanoparticles

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Abstract

A high-affinity monoclonal antibody (mAb) has been prepared and separately a gold nanoparticle (AuNP)-based and a near-infrared (NIR) fluorescence-based lateral flow immunoassay (LFA) developed for determination of 5-hydroxyflunixin residue in raw milk. The AuNP and IRDye® 800CW were used to label anti-5-hydroxyflunixin mAb to form the AuNP-mAb and NIR dye-mAb conjugates, respectively. Quantitative determination of 5-hydroxyflunixin was achieved by imaging the optical or fluorescence intensity of the AuNP-mAb and NIR dye-mAb captured on the test line. As a result, the detection limits of the AuNP-based LFA and NIR dye-based LFA were 0.82 and 0.073 ng/mL in raw milk, respectively. The considerable improvement on assay sensitivity of the NIR-based LFA can be attributed to the lower background and less antibody consumption per test than that of the AuNP-based LFA. The spiking experiment by the NIR-based LFA yielded 85.7–112.6% recovery with a relative standard deviation below 14%, indicating that it has satisfactory assay accuracy and precision. Furthermore, the analytical results of actual samples by the NIR dye-based LFA were consistent with that by instrumental analysis. Therefore, these results demonstrated that the NIR dye is an ideal alternative label to the conventional AuNP for the development of LFA for veterinary drugs in animal-origin food.

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Acknowledgments

We thank LetPub (www.letpub.com) for its linguistic assistance during the preparation of this manuscript.

Funding

This work is financially supported by Beijing Advanced Innovation Center for Food Nutrition and Human Health and the Chinese Universities Scientific Fund (2017304010328).

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Authors and Affiliations

  1. State Key Laboratory of Animal Nutrition, and Beijing Advanced Innovation Center for Food Nutrition and Human Health, China Agricultural University, Beijing, 100193, China

    Ruiqi Fan, Wanjun Zhang, Yongpeng Jin & Yiqiang Chen

  2. NBGen Co Ltd., Building 7, Bodaxing Industry Park, BDA, Beijing, 101111, China

    Rongmao Zhao & Chunjiang Yang

  3. Department of Cancer Genetics and Epigenetics, City of Hope National Medical Center, 1500 East Duarte Road, Duarte, CA, 91010, USA

    Qian Chen

  4. Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, 32306, USA

    Lidong He

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  1. Ruiqi Fan
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Correspondence to Yiqiang Chen.

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Fan, R., Zhang, W., Jin, Y. et al. Lateral flow immunoassay for 5-hydroxyflunixin based on near-infrared fluorescence molecule as an alternative label to gold nanoparticles. Microchim Acta 187, 368 (2020). https://doi.org/10.1007/s00604-020-04338-z

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