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Characterizing aptamer interaction with the oncolytic virus VV-GMCSF-Lact / M. A. Dymova, D. O. Malysheva, V. K. Popova [et al.] // Molecules. - 2024. - Vol. 29, Is. 4. - Ст. 848, DOI 10.3390/molecules29040848. - Cited References: 46. - This study was supported by the Russian Science Foundation grant No. 22-64-00041, available online: https://rscf.ru/en/project/22-64-00041/ (accessed on 6 February 2024). This work was supported by the Russian state-funded project for ICBFM SB RAS (grant number 121030200173-6) . - ISSN 1420-3049
Кл.слова (ненормированные):
aptamer -- oncolytic virus -- glioma -- dynamic light scattering -- microscale thermophoresis
Аннотация: Aptamers are currently being investigated for their potential to improve virotherapy. They offer several advantages, including the ability to prevent the aggregation of viral particles, enhance target specificity, and protect against the neutralizing effects of antibodies. The purpose of this study was to comprehensively investigate an aptamer capable of enhancing virotherapy. This involved characterizing the previously selected aptamer for vaccinia virus (VACV), evaluating the aggregation and molecular interaction of the optimized aptamers with the recombinant oncolytic virus VV-GMCSF-Lact, and estimating their immunoshielding properties in the presence of human blood serum. We chose one optimized aptamer, NV14t_56, with the highest affinity to the virus from the pool of several truncated aptamers and built its 3D model. The NV14t_56 remained stable in human blood serum for 1 h and bound to VV-GMCSF-Lact in the micromolar range (Kd ≈ 0.35 μM). Based on dynamic light scattering data, it has been demonstrated that aptamers surround viral particles and inhibit aggregate formation. In the presence of serum, the hydrodynamic diameter (by intensity) of the aptamer–virus complex did not change. Microscale thermophoresis (MST) experiments showed that NV14t_56 binds with virus (EC50 = 1.487 × 109 PFU/mL). The analysis of the amplitudes of MST curves reveals that the components of the serum bind to the aptamer–virus complex without disrupting it. In vitro experiments demonstrated the efficacy of VV-GMCSF-Lact in conjunction with the aptamer when exposed to human blood serum in the absence of neutralizing antibodies (Nabs). Thus, NV14t_56 has the ability to inhibit virus aggregation, allowing VV-GMCSF-Lact to maintain its effectiveness throughout the storage period and subsequent use. When employing aptamers as protective agents for oncolytic viruses, the presence of neutralizing antibodies should be taken into account.

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Держатели документа:
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev av. 8, 630090 Novosibirsk, Russia
Department of Natural Sciences, Novosibirsk State University, Pirogova str. 1, 630090 Novosibirsk, Russia
State Research Center of Virology and Biotechnology “Vector”, 630559 Koltsovo, Russia
Laboratory for Biomolecular and Medical Technologies, Krasnoyarsk State Medical University Named after Prof. V.F. Voyno-Yasenetsky, Partizana Zheleznyaka str. 1, 660022 Krasnoyarsk, Russia
Federal Research Center KSC SB RAS, 50 Akademgorodok, 660036 Krasnoyarsk, Russia
Kirensky Institute of Physics, 50/38 Akademgorodok, 660012 Krasnoyarsk, Russia

Доп.точки доступа:
Dymova, M. A.; Malysheva, D. O.; Popova, V. K.; Dmitrienko, E. V.; Endutkin, A. V.; Drokov, D. V.; Mukhanov, V. S.; Byvakina, A. A.; Kochneva, G. V.; Artyushenko, P. V.; Shchugoreva, I. A.; Rogova, A. V.; Tomilin, F. N.; Томилин, Феликс Николаевич; Kichkailo, A. S.; Richter, V. A.; Kuligina, E. V.
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