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Full‐length G glycoprotein directly extracted from rabies virus with detergent and then stabilized by amphipols in liquid and freeze‐dried forms
Pathogen surface antigens are at the forefront of the viral strategy when invading host organisms. These antigens, including membrane proteins (MPs), are broadly targeted by the host immune response. Obtaining these MPs in a soluble and stable form constitutes a real challenge, regardless of the app...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9291542/ https://www.ncbi.nlm.nih.gov/pubmed/34297405 http://dx.doi.org/10.1002/bit.27900 |
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author | Clénet, Didier Clavier, Léna Strobbe, Benoît Le Bon, Christel Zoonens, Manuela Saulnier, Aure |
author_facet | Clénet, Didier Clavier, Léna Strobbe, Benoît Le Bon, Christel Zoonens, Manuela Saulnier, Aure |
author_sort | Clénet, Didier |
collection | PubMed |
description | Pathogen surface antigens are at the forefront of the viral strategy when invading host organisms. These antigens, including membrane proteins (MPs), are broadly targeted by the host immune response. Obtaining these MPs in a soluble and stable form constitutes a real challenge, regardless of the application purposes (e.g. quantification/characterization assays, diagnosis, and preventive and curative strategies). A rapid process to obtain a native‐like antigen by solubilization of a full‐length MP directly from a pathogen is reported herein. Rabies virus (RABV) was used as a model for this demonstration and its full‐length G glycoprotein (RABV‐G) was stabilized with amphipathic polymers, named amphipols (APols). The stability of RABV‐G trapped in APol A8‐35 (RABV‐G/A8‐35) was evaluated under different stress conditions (temperature, agitation, and light exposure). RABV‐G/A8‐35 in liquid form exhibited higher unfolding temperature (+6°C) than in detergent and was demonstrated to be antigenically stable over 1 month at 5°C and 25°C. Kinetic modeling of antigenicity data predicted antigenic stability of RABV‐G/A8‐35 in a solution of up to 1 year at 5°C. The RABV‐G/A8‐35 complex formulated in an optimized buffer composition and subsequently freeze‐dried displayed long‐term stability for 2‐years at 5, 25, and 37°C. This study reports for the first time that a natural full‐length MP extracted from a virus, complexed to APols and subsequently freeze‐dried, displayed long‐term antigenic stability, without requiring storage under refrigerated conditions. |
format | Online Article Text |
id | pubmed-9291542 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-92915422022-07-20 Full‐length G glycoprotein directly extracted from rabies virus with detergent and then stabilized by amphipols in liquid and freeze‐dried forms Clénet, Didier Clavier, Léna Strobbe, Benoît Le Bon, Christel Zoonens, Manuela Saulnier, Aure Biotechnol Bioeng ARTICLES Pathogen surface antigens are at the forefront of the viral strategy when invading host organisms. These antigens, including membrane proteins (MPs), are broadly targeted by the host immune response. Obtaining these MPs in a soluble and stable form constitutes a real challenge, regardless of the application purposes (e.g. quantification/characterization assays, diagnosis, and preventive and curative strategies). A rapid process to obtain a native‐like antigen by solubilization of a full‐length MP directly from a pathogen is reported herein. Rabies virus (RABV) was used as a model for this demonstration and its full‐length G glycoprotein (RABV‐G) was stabilized with amphipathic polymers, named amphipols (APols). The stability of RABV‐G trapped in APol A8‐35 (RABV‐G/A8‐35) was evaluated under different stress conditions (temperature, agitation, and light exposure). RABV‐G/A8‐35 in liquid form exhibited higher unfolding temperature (+6°C) than in detergent and was demonstrated to be antigenically stable over 1 month at 5°C and 25°C. Kinetic modeling of antigenicity data predicted antigenic stability of RABV‐G/A8‐35 in a solution of up to 1 year at 5°C. The RABV‐G/A8‐35 complex formulated in an optimized buffer composition and subsequently freeze‐dried displayed long‐term stability for 2‐years at 5, 25, and 37°C. This study reports for the first time that a natural full‐length MP extracted from a virus, complexed to APols and subsequently freeze‐dried, displayed long‐term antigenic stability, without requiring storage under refrigerated conditions. John Wiley and Sons Inc. 2021-08-05 2021-11 /pmc/articles/PMC9291542/ /pubmed/34297405 http://dx.doi.org/10.1002/bit.27900 Text en © 2021 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals LLC https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | ARTICLES Clénet, Didier Clavier, Léna Strobbe, Benoît Le Bon, Christel Zoonens, Manuela Saulnier, Aure Full‐length G glycoprotein directly extracted from rabies virus with detergent and then stabilized by amphipols in liquid and freeze‐dried forms |
title | Full‐length G glycoprotein directly extracted from rabies virus with detergent and then stabilized by amphipols in liquid and freeze‐dried forms |
title_full | Full‐length G glycoprotein directly extracted from rabies virus with detergent and then stabilized by amphipols in liquid and freeze‐dried forms |
title_fullStr | Full‐length G glycoprotein directly extracted from rabies virus with detergent and then stabilized by amphipols in liquid and freeze‐dried forms |
title_full_unstemmed | Full‐length G glycoprotein directly extracted from rabies virus with detergent and then stabilized by amphipols in liquid and freeze‐dried forms |
title_short | Full‐length G glycoprotein directly extracted from rabies virus with detergent and then stabilized by amphipols in liquid and freeze‐dried forms |
title_sort | full‐length g glycoprotein directly extracted from rabies virus with detergent and then stabilized by amphipols in liquid and freeze‐dried forms |
topic | ARTICLES |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9291542/ https://www.ncbi.nlm.nih.gov/pubmed/34297405 http://dx.doi.org/10.1002/bit.27900 |
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