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Development of a biosafe ELISA-based platform for assessing immunogenicity in the production of an inactivated whole-virion coronavirus vaccine
- Authors: Danilov D.V.1, Shmeleva O.A.1, Lunin A.S.1, Kozlovskaya L.I.1, Piniaeva A.N.1, Shishova A.A.1
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Affiliations:
- Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
- Issue: Vol 22, No 2 (2022)
- Pages: 163-169
- Section: Conference proceedings
- URL: https://journals.eco-vector.com/MAJ/article/view/108717
- DOI: https://doi.org/10.17816/MAJ108717
- ID: 108717
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Abstract
BACKGROUND: SARS-CoV-2 vaccine immunogenicity is evaluated in neutralization test with live virus. It is performed in a biosafety level 3 zone because requires live virus stage. Therefore, control laboratories should be certified for this class of work. The development of technology based on enzyme-linked immunosorbent assay as an analogue of the neutralization reaction makes it possible to create an immunobiological product in a shorter time and in conditions without special requirements for control laboratories.
AIM: Development of an enzyme-linked immunosorbent assay for assessing SARS-CoV-2 vaccine immunogenicity by measuring neutralizing antibodies production in immunized animals.
MATERIALS AND METHODS: Recombinant receptor-binding domain fused to a С-terminal hexahistidine sequence was produced in Escherichia coli cells and purified via metal-affinity chromatography on WorkBeads NiMAC (Bio-Works). Purified protein was used in enzyme-linked immunosorbent assay as an antigen for sorption. The sera of mice immunized with the vaccine preparation were tested for neutralizing activity against the SARS-CoV-2, as well as in the developed enzyme-linked immunosorbent assay.
RESULTS: Sera with high neutralizing titers showed a high degree of binding to recombinant receptor-binding domain fused to a С-terminal hexahistidine sequence in enzyme-linked immunosorbent assay, while sera from non-immunized animals or sera with neutralization titers less than 1:8 were not reactive in enzyme-linked immunosorbent assay. The Spearman and Pearson correlation coefficients for neutralization test titers and optical density in enzyme-linked immunosorbent assay were 0.759 and 0.76, respectively. The developed assay can be used as a semi-quantitative method for assessing the immunogenicity of a vaccine against coronavirus infection.
CONCLUSIONS: The developed platform makes it possible to reliably assess the immunogenicity of an inactivated coronavirus vaccine under conditions that do not require a high biosafety conditions.
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About the authors
Dmitry V. Danilov
Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Author for correspondence.
Email: danilov_dv@chumakovs.su
ORCID iD: 0000-0002-5539-1112
Technologist of Analytical Method Development and Validation Team
Russian Federation, MoscowOlga A. Shmeleva
Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: olya.shmeleva.2000@mail.ru
Lab Assistant of Analytical Method Development and Validation Team
Russian Federation, MoscowAlexander S. Lunin
Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: lunin_as@chumakovs.su
Virologist of Preclinical Research Department
Russian Federation, MoscowLiubov I. Kozlovskaya
Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: kozlovskaya_li@chumakovs.su
ORCID iD: 0000-0002-3029-1035
SPIN-code: 6865-9260
Scopus Author ID: 12646876000
ResearcherId: E-2368-2014
Cand. Sci. (Biol.), Head, Department of Emerging and Reemerging Infections with Pandemic Potential
Russian Federation, MoscowAnastasia N. Piniaeva
Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: an_Piniaeva@chumakovs.su
ORCID iD: 0000-0001-5381-2393
Scopus Author ID: 57218545661
Head of Division of Development and Integration of Innovative and Semi-industrial Technologies
Russian Federation, MoscowAnna A. Shishova
Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: aa_shishova@chumakovs.su
ORCID iD: 0000-0002-5907-0615
Research Associate, Laboratory of BioChemistry
Russian Federation, MoscowReferences
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