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Volume 15
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Article

Development of a Multiplex Lateral Flow Immunoassay for the Detection of Antibiotics in Milk Utilizing Lyophilized Gold Nanoparticle Conjugates

by
Ivan V. Maksin
1,2,*,
Azhar Kuandykova
2,
Darya I. Polyakova
2,3,
Viktoriia A. Kesareva
2,3,
Timofei A. Luzyanin
1,
Vladislav S. Ivanov
2,
Evgeniia I. Simonova
2,4,
German A. Khunteev
2 and
Yuliya G. Kirillova
1
1
Department of Biotechnology and Industrial Pharmacy, Lomonosov Institute of Fine Chemical Technologies, MIREA–Russian Technological University, 86, Vernadsky Avenue, 119571 Moscow, Russia
2
Rapid Bio LLC, 42, bld. 1, Bolshoy Boulevard, 121205 Moscow, Russia
3
Institute of Pharmacy and Biotechnology, Peoples’ Friendship University of Russia named after Patrice Lumumba, 8, bld. 2, Miklukho-Maklaya, 117198 Moscow, Russia
4
Department of Veterinary Medicine, Agro–Technological Institute, Peoples’ Friendship University of Russia named after Patrice Lumumba, 8, bld. 2, Miklukho-Maklaya, 117198 Moscow, Russia
*
Author to whom correspondence should be addressed.
Biosensors 2025, 15(9), 592; https://doi.org/10.3390/bios15090592
Submission received: 18 July 2025 / Revised: 1 September 2025 / Accepted: 3 September 2025 / Published: 9 September 2025
(This article belongs to the Special Issue Immunosensors: Design and Applications)
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Abstract

Lateral flow immunoassays (LFAs) are widely recognized as a powerful and versatile analytical platform. Nevertheless, the development of multiplex formats remains a distinct challenge. The aim of this study was to develop a multiplex LFA using gold nanoparticles (GNPs) as a label, selected for their ease of synthesis and functionalization with biomolecules. We provide practical recommendations regarding protein–hapten synthesis, membrane selection, application buffer composition, and methods to improve the long-term stability of the freeze-dried gold conjugate. The developed assay shows good tolerance to high-fat milk, stability at elevated temperatures, and promising sensitivity, with visual detection limits of 4–100 ng/mL for β-lactams, 1–10 ng/mL for tetracyclines, 50 ng/mL for streptomycin, and 0.3 ng/mL for chloramphenicol.
Keywords: immunosensor; rapid test; gold conjugate; freeze-drying; antibody; optimization; lateral flow immunoassay immunosensor; rapid test; gold conjugate; freeze-drying; antibody; optimization; lateral flow immunoassay

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MDPI and ACS Style

Maksin, I.V.; Kuandykova, A.; Polyakova, D.I.; Kesareva, V.A.; Luzyanin, T.A.; Ivanov, V.S.; Simonova, E.I.; Khunteev, G.A.; Kirillova, Y.G. Development of a Multiplex Lateral Flow Immunoassay for the Detection of Antibiotics in Milk Utilizing Lyophilized Gold Nanoparticle Conjugates. Biosensors 2025, 15, 592. https://doi.org/10.3390/bios15090592

AMA Style

Maksin IV, Kuandykova A, Polyakova DI, Kesareva VA, Luzyanin TA, Ivanov VS, Simonova EI, Khunteev GA, Kirillova YG. Development of a Multiplex Lateral Flow Immunoassay for the Detection of Antibiotics in Milk Utilizing Lyophilized Gold Nanoparticle Conjugates. Biosensors. 2025; 15(9):592. https://doi.org/10.3390/bios15090592

Chicago/Turabian Style

Maksin, Ivan V., Azhar Kuandykova, Darya I. Polyakova, Viktoriia A. Kesareva, Timofei A. Luzyanin, Vladislav S. Ivanov, Evgeniia I. Simonova, German A. Khunteev, and Yuliya G. Kirillova. 2025. "Development of a Multiplex Lateral Flow Immunoassay for the Detection of Antibiotics in Milk Utilizing Lyophilized Gold Nanoparticle Conjugates" Biosensors 15, no. 9: 592. https://doi.org/10.3390/bios15090592

APA Style

Maksin, I. V., Kuandykova, A., Polyakova, D. I., Kesareva, V. A., Luzyanin, T. A., Ivanov, V. S., Simonova, E. I., Khunteev, G. A., & Kirillova, Y. G. (2025). Development of a Multiplex Lateral Flow Immunoassay for the Detection of Antibiotics in Milk Utilizing Lyophilized Gold Nanoparticle Conjugates. Biosensors, 15(9), 592. https://doi.org/10.3390/bios15090592

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