Production and purification of recombinant proteins VP2 and VP3 of the <i>Alongshan</i> virus of the <i>Jingmenvirus</i> group and evaluation of their immunochemical properties
Introduction. Alongshan virus is a representative of the unclassified group of Jingmenviruses (Flaviviridae), which is detected in Ixodes persulcatus, Ixodes ricinus ticks and various mosquito species in Russia, China, Finland and France. Unlike traditional orthoflaviviruses, the Alongshan virus gen...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | Russian |
| Published: |
Central Research Institute for Epidemiology
2025-05-01
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| Series: | Журнал микробиологии, эпидемиологии и иммунобиологии |
| Subjects: | |
| Online Access: | https://microbiol.crie.ru/jour/article/viewFile/18829/1593 |
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| Summary: | Introduction. Alongshan virus is a representative of the unclassified group of Jingmenviruses (Flaviviridae), which is detected in Ixodes persulcatus, Ixodes ricinus ticks and various mosquito species in Russia, China, Finland and France. Unlike traditional orthoflaviviruses, the Alongshan virus genome is represented by 4 positive-sense RNA segments. The first and third segments of the genome encode proteins homologous to proteins of the replicative machinery of orthoflaviviruses, the remaining segments encode putative structural proteins that have no known homologues: segment 2 — VP1a (envelope protein), VP1b and NuORF; segment 4 — VP2 (capsid) and VP3 (membrane). Human cases of Alongshan virus-associated disease have been described.
The aim of this study is to develop a system for expression and purification of recombinant VP2 and hydrophilic site VP3 proteins to test their antigenic properties.
Materials and methods. Miass527 strain of Alongshan virus was used to produce hyperimmune mouse sera and recombinant proteins in a bacterial expression system. Bioinformatic analysis of sequences encoding target proteins and genetically engineered cloning were carried out in this study. Western blotting and enzyme immunoassay (ELISA) were performed to control the results.
Results. Recombinant proteins of Alongshan virus have been used in a laboratory diagnostic test system to determine the presence of antibodies to the virus. The obtained recombinant VP2 protein is able to detect antibodies in all tested sera of infected mice, as well as antibodies in human sera both in Western blotting and in enzyme immunoassay. At the same time, antibodies to the recombinant region of VP3 protein were detected irregularly in antiviral immune sera.
Conclusion. The detection of antibodies to Alongshan virus in patients confirms the necessity for further investigation of this group of viruses. |
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| ISSN: | 0372-9311 2686-7613 |