Zhdanov Dmitry D, Shishparenok Anastasia N, Ivin Yury Y, Kovpak Anastasia A, Piniaeva Anastasia N, Levin Igor V, Budnik Sergei V, Shilov Oleg A, Churyukin Roman S, Agafonova Lubov E, Berezhnova Alina V, Shumyantseva Victoria V, Ishmukhametov Aydar A
Institute of Biomedical Chemistry, 10 Pogodinskaya str., 119121 Moscow, Russia.
Department of Biochemistry, People's Friendship University of Russia Named After Patrice Lumumba (RUDN University), 6 Miklukho-Maklaya str., 117198 Moscow, Russia.
Vaccines (Basel). 2025 Jul 31;13(8):824. doi: 10.3390/vaccines13080824.
Most antiviral vaccines are created by inactivating the virus using chemical methods. The inactivation and production of viral vaccine preparations after the irradiation of viruses with accelerated electrons has a number of significant advantages. Determining the integrity of the genome of the resulting viral particles is necessary to assess the quality and degree of inactivation after irradiation.
This work was performed on the Sabin 2 model polio virus. To determine the most sensitive and most radiation-resistant part, the polio virus genome was divided into 20 segments. After irradiation at temperatures of 25 °C, 2-8 °C, -20 °C, or -70 °C, the amplification intensity of these segments was measured in real time.
The best correlation between the amplification cycle and the irradiation dose at all temperatures was observed for segment 3D, left. Consequently, this section of the poliovirus genome is the least resistant to the action of accelerated electrons and is the most representative for determining genome integrity. The worst dependence was observed for the VP1 right section, which, therefore, cannot be used to determine genome integrity during inactivation. The electrochemical approach was also employed for a comparative assessment of viral RNA integrity before and after irradiation. An increase in the irradiation dose was accompanied by an increase in signals indicating the electrooxidation of RNA heterocyclic bases. The increase in peak current intensity of viral RNA electrochemical signals confirmed the breaking of viral RNA strands during irradiation. The shorter the RNA fragments, the greater the peak current intensities. In turn, this made the heterocyclic bases more accessible to electrooxidation on the electrode.
These results are necessary for characterizing the integrity of the viral genome for the purpose of creating of antiviral vaccines.
大多数抗病毒疫苗是通过化学方法使病毒失活而制备的。用加速电子照射病毒后制备病毒疫苗制剂具有许多显著优点。确定所得病毒颗粒基因组的完整性对于评估照射后的质量和失活程度是必要的。
本研究以Sabin 2型脊髓灰质炎病毒为模型。为了确定最敏感和最耐辐射的部分，将脊髓灰质炎病毒基因组分为20个片段。在25℃、2 - 8℃、-20℃或-70℃温度下照射后，实时测量这些片段的扩增强度。
在所有温度下，观察到3D片段左侧的扩增循环与照射剂量之间的相关性最佳。因此，脊髓灰质炎病毒基因组的这一部分对加速电子的作用最不耐受，是确定基因组完整性最具代表性的部分。观察到VP1右侧片段的相关性最差，因此，在失活过程中不能用它来确定基因组完整性。还采用电化学方法对比评估照射前后病毒RNA的完整性。照射剂量增加伴随着表明RNA杂环碱基电氧化的信号增加。病毒RNA电化学信号的峰值电流强度增加证实了照射过程中病毒RNA链的断裂。RNA片段越短，峰值电流强度越大。相应地，这使得杂环碱基在电极上更易于发生电氧化。
这些结果对于表征病毒基因组的完整性以制备抗病毒疫苗具有重要意义。
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