Study of the influence of nucleotide sequence optimization of the target gene on the immunogenicity of a recombinant adenovirus vector

Zubkova O.V.; Ozharovskaia T.A.; Popova O.; Korobova E.V.; Goldovskaya P.P.; Zrelkin D.I.; Shcherbinin D.N.; Shcheblyakov D.V.

doi:https://doi.org/10.17116/molgen20244204143

Zubkova O.V.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0001-7893-8419

Ozharovskaia T.A.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0001-7147-1553

Popova O.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0003-3248-1227

Korobova E.V.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya;
MIREA — Russian Technological University

ORCID: 0009-0000-4511-0164

Goldovskaya P.P.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya;
MIREA — Russian Technological University

ORCID: 0009-0000-1965-0482

Zrelkin D.I.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0003-0899-8357

Shcherbinin D.N.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0002-8518-1669

Shcheblyakov D.V.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0002-1289-3411

Study of the influence of nucleotide sequence optimization of the target gene on the immunogenicity of a recombinant adenovirus vector

Authors:

Zubkova O.V., Ozharovskaia T.A., Popova O., Korobova E.V., Goldovskaya P.P., Zrelkin D.I., Shcherbinin D.N., Shcheblyakov D.V.

More about the authors

Journal: Molecular Genetics, Microbiology and Virology. 2024;42(4): 43‑48

DOI: 10.17116/molgen20244204143

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To cite this article:

Zubkova OV, Ozharovskaia TA, Popova O, et al. . Study of the influence of nucleotide sequence optimization of the target gene on the immunogenicity of a recombinant adenovirus vector. Molecular Genetics, Microbiology and Virology. 2024;42(4):43‑48. (In Russ.)
https://doi.org/10.17116/molgen20244204143

Подписка 2026 Молекулярная генетика, микробиология и вирусология (Molecular Genetics, Microbiology and Virology)

Использование инфографики авторами научных работ

THE PURPOSE OF THE STUDY

Was to compare the immunogenicity of recombinant adenoviruses expressing the S protein gene of the SARS-CoV-2 virus variant Omicron (BA.5.3.1) with an optimized nucleotide sequence (Ad5-S-BA5opt) or non-optimized (Ad5-S-BA5).

MATERIAL AND METHODS

Recombinant adenoviruses were grown on HEK293 cell culture; purification was carried out by ultracentrifugation in a stepwise density gradient of CsCl. The resulting adenovirus samples were characterized by authenticity (by real-time PCR (RT-PCR) and immunoblotting), the number of viral particles (using spectrophotometry) and infectious titer (using the end-point dilution (TCD₅₀) method). The titer of RBD-specific IgG antibodies in an animal model was assessed by enzyme-linked immunosorbent assay (ELISA).

RESULTS

Based on the results of studies in an animal model, a vector expressing the optimized gene (Ad5-S-BA5opt) led to the formation of higher levels of IgG-specific antibodies compared to a similar vector containing the wild-type protein gene. Particularly interesting is the fact that an approach aimed solely at optimizing the sequence of the target gene, and not other components of the vector, can make significant changes in the immunogenicity of the vector.

CONCLUSION

The obtained results are important for the development of new, more effective strategies for creating vaccines and gene therapy drugs. Optimization of the nucleotide sequence can lead to an improved mRNA stability and a significant increase in the expression of the target gene, and as a result, a significant increase in the protective properties of the candidate drug. This method can be assumed to become promising for the development of vaccines against viruses containing low-immunogenic antigens.

Keywords:

Recombinant adenoviral vectors

vaccines

S protein of SARS-CoV-2

codon optimization

codon usage

immunogenicity

Authors:

Zubkova O.V.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0001-7893-8419

Ozharovskaia T.A.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0001-7147-1553

Popova O.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0003-3248-1227

Korobova E.V.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya;
MIREA — Russian Technological University

ORCID: 0009-0000-4511-0164

Goldovskaya P.P.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya;
MIREA — Russian Technological University

ORCID: 0009-0000-1965-0482

Zrelkin D.I.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0003-0899-8357

Shcherbinin D.N.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0002-8518-1669

Shcheblyakov D.V.

National Research Center for Epidemiology and Microbiology named after the honorary academician N.F. Gamaleya

ORCID: 0000-0002-1289-3411

Received:

11.03.2024

Accepted:

25.10.2024

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