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Application of A29L Protein Specific Monoclonal Antibodies A-A29L_MPoxV for Monkeypox Diagnosis

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Abstract

The spread of the disease caused by monkeypox virus (MPox) since 2022 has shown the urgency of developing countermeasures. The development of modern methods of clinical laboratory diagnostics of MPox contributes to this. Enzyme-linked immunosorbent assay (ELISA) is an accessible and sensitive platform for developing diagnostic tools. Detection of MPox antigens using ELISA kits based on monoclonal antibodies (MAbs) is promising due to the quick time of analysis and minimal requirements for sample preparation. We have developed and deposited two strains of Escherichia coli that produce recombinant proteins. Mice were immunized with the AgPOX protein, which contains unique antigenic sequences of MPox. The Trx + A29 protein for selecting MAb producers includes the original amino acid sequence A29L. The absence of antibody crossover to Trx protein and native preparations of variola virus and vaccinia virus tested by ELISA. As a result of hybridization of splenocytes from immunized mice, MAb producers were obtained. Fifteen MAb-producing hybridomas were selected based on ELISA results with three specific MPox antigens and three nonspecific ones. Three hybridomas were selected for deposit according to the productivity criteria. The possibility of detection by means of its MAbs of the native MPox antigen at various concentrations was tested and method sensitivity was determined. The MAbs a-A29L_MPoxV of three hybridomas detected the native antigen MPox at a concentration of 102 PFU/mL. It is likely that the method is even more sensitive when selecting analysis conditions. Based on labeled MAbs a-A29L_MPoxV, it is possible to develop a sensitive and specific indirect two-step ELISA kit for immunodiagnostics of MPox.

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ACKNOWLEDGMENTS

The authors are grateful to Andrei Leonidovich Matveev, Senior Researcher of the Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of the Russian Academy of Sciences, for his invaluable contribution to the creation of hybridomas and editing the article.

Funding

The study was supported by the state order GZ-34/21 to the State Scientific Center of Virology and Biotechnology “VECTOR,” project no. 121033100069-4.

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Authors and Affiliations

  1. State Scientific Center of Virology and Biotechnology “VECTOR”, 630559, Koltsovo, Novosibirsk oblast, Russia

    S. A. Pyankov, I. S. Shulgina, A. V. Rybel, A. Z. Maksyutov, V. Yu. Tyurin, I. A. Drachkova, T. V. Tregubchak, T. V. Bauer, A. S. Ovchinnikova, D. A. Odnoshevskiy, A. S. Kabanov, S. A. Bodnev, O. V. Pyankov & A. P. Agafonov

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  1. S. A. Pyankov
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  2. I. S. Shulgina
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  4. A. Z. Maksyutov
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  7. T. V. Tregubchak
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  8. T. V. Bauer
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  9. A. S. Ovchinnikova
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  11. A. S. Kabanov
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  13. O. V. Pyankov
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  14. A. P. Agafonov
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Correspondence to S. A. Pyankov.

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The authors of this work declare that they have no conflicts of interest.

ETHICS APPROVAL AND CONSENT TO PARTICIPATE

Animal-related experiments were conducted in accordance with the NIH Guidelines for the care and use of laboratory animals (http://oacu.od.nih.gov/regs/index.htm). Animal protocols were approved by the Bioethics Committee of the State Scientific Center of Virology and Biotechnology “VECTOR.”

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Translated by A. Kazantseva

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Pyankov, S.A., Shulgina, I.S., Rybel, A.V. et al. Application of A29L Protein Specific Monoclonal Antibodies A-A29L_MPoxV for Monkeypox Diagnosis. Mol. Genet. Microbiol. Virol. 38, 242–248 (2023). https://doi.org/10.3103/S0891416823040067

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  • DOI: https://doi.org/10.3103/S0891416823040067

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