Development and evaluation of a recombinant monoclonal human antibody with virus-neutralizing activity against the F glycoprotein of respiratory syncytial virus
Introduction. Respiratory syncytial virus (RSV) is the most common pathogen causing lower respiratory tract infections in children. RSV also poses a serious threat to the elderly and immunocompromised patients. Developing a therapy based on recombinant human antibodies to block the RSV fusion (F) gl...
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| Language: | Russian |
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Central Research Institute for Epidemiology
2024-12-01
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| Series: | Журнал микробиологии, эпидемиологии и иммунобиологии |
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| Online Access: | https://microbiol.crie.ru/jour/article/viewFile/18697/1548 |
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| author | Sergey A. Klotchenko Ekaterina A. Romanovskaya-Romanko Marina A. Plotnikova Anastasia A. Pulkina Aram A. Shaldzhyan Dmitry S. Balabashin Victoria A. Toporova Teimur K. Aliev Natalia E. Gyulikhandanova Dmitry A. Lioznov |
| author_facet | Sergey A. Klotchenko Ekaterina A. Romanovskaya-Romanko Marina A. Plotnikova Anastasia A. Pulkina Aram A. Shaldzhyan Dmitry S. Balabashin Victoria A. Toporova Teimur K. Aliev Natalia E. Gyulikhandanova Dmitry A. Lioznov |
| author_sort | Sergey A. Klotchenko |
| collection | DOAJ |
| description | Introduction. Respiratory syncytial virus (RSV) is the most common pathogen causing lower respiratory tract infections in children. RSV also poses a serious threat to the elderly and immunocompromised patients. Developing a therapy based on recombinant human antibodies to block the RSV fusion (F) glycoprotein is urgent to reduce the incidence of RSV infections and prevent associated complications.
Aim. To design plasmid vectors for efficient production of the recombinant monoclonal antibody FM1 in a eukaryotic expression system targeting the RSV fusion (F) glycoprotein and to evaluate its activity against RSV subtypes A and B in vitro.
Materials and methods. Constructs encoding the recombinant antibody FM1 were designed using genetic engineering. Recombinant antibodies were produced in the CHO-K1 cell line through transient expression. Antibody specimens were purified from the culture supernatant using affinity chromatography, with a modified protein A as the ligand. The virus-neutralizing activity of the antibody was evaluated in a microneutralization assay using several RSV strains on a Vero cell monolayer culture.
Results. We developed a two-plasmid vector system to produce the recombinant FM1 antibody targeting the RSV F glycoprotein, using CHO cells as transient producers. The antibody was successfully produced, purified, and characterized, with its biological activity confirmed. The FM1 antibody demonstrated enhanced virus-neutralizing activity against reference and seasonal RSV strains of subtypes A and B compared to the control drug palivizumab.
Conclusion. A recombinant FM1 antibody-based drug could address the import substitution challenge for protective measures against RSV infection. The authors are currently developing a stable FM1 producer clone with high productivity and viability and investigating the therapeutic efficacy of this antibody in a sublethal RSV infection mouse model. |
| format | Article |
| id | doaj-art-46ea58ef7d2a4c4db55693a879e5c12f |
| institution | Kabale University |
| issn | 0372-9311 2686-7613 |
| language | Russian |
| publishDate | 2024-12-01 |
| publisher | Central Research Institute for Epidemiology |
| record_format | Article |
| series | Журнал микробиологии, эпидемиологии и иммунобиологии |
| spelling | doaj-art-46ea58ef7d2a4c4db55693a879e5c12f2025-02-06T21:11:31ZrusCentral Research Institute for EpidemiologyЖурнал микробиологии, эпидемиологии и иммунобиологии0372-93112686-76132024-12-01101673574710.36233/0372-9311-6112795Development and evaluation of a recombinant monoclonal human antibody with virus-neutralizing activity against the F glycoprotein of respiratory syncytial virusSergey A. Klotchenko0https://orcid.org/0000-0003-0289-6560Ekaterina A. Romanovskaya-Romanko1https://orcid.org/0000-0001-7560-398XMarina A. Plotnikova2https://orcid.org/0000-0001-8196-3156Anastasia A. Pulkina3https://orcid.org/0000-0001-8609-8093Aram A. Shaldzhyan4https://orcid.org/0000-0002-8646-6252Dmitry S. Balabashin5https://orcid.org/0000-0002-7627-0600Victoria A. Toporova6https://orcid.org/0000-0002-7450-7096Teimur K. Aliev7https://orcid.org/0000-0002-1753-9614Natalia E. Gyulikhandanova8https://orcid.org/0000-0001-6907-0144Dmitry A. Lioznov9https://orcid.org/0000-0003-3643-7354Smorodintsev Research Institute of InfluenzaSmorodintsev Research Institute of InfluenzaSmorodintsev Research Institute of InfluenzaSmorodintsev Research Institute of InfluenzaSmorodintsev Research Institute of InfluenzaShemyakin–Ovchinnikov Institute of bioorganic chemistrySmorodintsev Research Institute of InfluenzaShemyakin–Ovchinnikov Institute of bioorganic chemistrySmorodintsev Research Institute of InfluenzaSmorodintsev Research Institute of InfluenzaIntroduction. Respiratory syncytial virus (RSV) is the most common pathogen causing lower respiratory tract infections in children. RSV also poses a serious threat to the elderly and immunocompromised patients. Developing a therapy based on recombinant human antibodies to block the RSV fusion (F) glycoprotein is urgent to reduce the incidence of RSV infections and prevent associated complications. Aim. To design plasmid vectors for efficient production of the recombinant monoclonal antibody FM1 in a eukaryotic expression system targeting the RSV fusion (F) glycoprotein and to evaluate its activity against RSV subtypes A and B in vitro. Materials and methods. Constructs encoding the recombinant antibody FM1 were designed using genetic engineering. Recombinant antibodies were produced in the CHO-K1 cell line through transient expression. Antibody specimens were purified from the culture supernatant using affinity chromatography, with a modified protein A as the ligand. The virus-neutralizing activity of the antibody was evaluated in a microneutralization assay using several RSV strains on a Vero cell monolayer culture. Results. We developed a two-plasmid vector system to produce the recombinant FM1 antibody targeting the RSV F glycoprotein, using CHO cells as transient producers. The antibody was successfully produced, purified, and characterized, with its biological activity confirmed. The FM1 antibody demonstrated enhanced virus-neutralizing activity against reference and seasonal RSV strains of subtypes A and B compared to the control drug palivizumab. Conclusion. A recombinant FM1 antibody-based drug could address the import substitution challenge for protective measures against RSV infection. The authors are currently developing a stable FM1 producer clone with high productivity and viability and investigating the therapeutic efficacy of this antibody in a sublethal RSV infection mouse model.https://microbiol.crie.ru/jour/article/viewFile/18697/1548therapeutic monoclonal antibodiesrespiratory syncytial virusfusion (f) glycoproteinneutralization assaycho cell line for recombinant antibodies productionvero cells |
| spellingShingle | Sergey A. Klotchenko Ekaterina A. Romanovskaya-Romanko Marina A. Plotnikova Anastasia A. Pulkina Aram A. Shaldzhyan Dmitry S. Balabashin Victoria A. Toporova Teimur K. Aliev Natalia E. Gyulikhandanova Dmitry A. Lioznov Development and evaluation of a recombinant monoclonal human antibody with virus-neutralizing activity against the F glycoprotein of respiratory syncytial virus Журнал микробиологии, эпидемиологии и иммунобиологии therapeutic monoclonal antibodies respiratory syncytial virus fusion (f) glycoprotein neutralization assay cho cell line for recombinant antibodies production vero cells |
| title | Development and evaluation of a recombinant monoclonal human antibody with virus-neutralizing activity against the F glycoprotein of respiratory syncytial virus |
| title_full | Development and evaluation of a recombinant monoclonal human antibody with virus-neutralizing activity against the F glycoprotein of respiratory syncytial virus |
| title_fullStr | Development and evaluation of a recombinant monoclonal human antibody with virus-neutralizing activity against the F glycoprotein of respiratory syncytial virus |
| title_full_unstemmed | Development and evaluation of a recombinant monoclonal human antibody with virus-neutralizing activity against the F glycoprotein of respiratory syncytial virus |
| title_short | Development and evaluation of a recombinant monoclonal human antibody with virus-neutralizing activity against the F glycoprotein of respiratory syncytial virus |
| title_sort | development and evaluation of a recombinant monoclonal human antibody with virus neutralizing activity against the f glycoprotein of respiratory syncytial virus |
| topic | therapeutic monoclonal antibodies respiratory syncytial virus fusion (f) glycoprotein neutralization assay cho cell line for recombinant antibodies production vero cells |
| url | https://microbiol.crie.ru/jour/article/viewFile/18697/1548 |
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