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Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic Coupling
We have developed a new binary epitope-presenting CVP platform based on bamboo mosaic virus (BaMV) by using the sortase A (SrtA)-mediated ligation technology. The reconstructed BaMV genome harbors two modifications: 1) a coat protein (CP) with N-terminal extension of the tobacco etch virus (TEV) pro...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8551676/ https://www.ncbi.nlm.nih.gov/pubmed/34721406 http://dx.doi.org/10.3389/fimmu.2021.739837 |
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author | Yang, Ming-Hao Hu, Chung-Chi Wong, Chi-Hzeng Liang, Jian-Jong Ko, Hui-Ying He, Meng-Hsun Lin, Yi-Ling Lin, Na-Sheng Hsu, Yau-Heiu |
author_facet | Yang, Ming-Hao Hu, Chung-Chi Wong, Chi-Hzeng Liang, Jian-Jong Ko, Hui-Ying He, Meng-Hsun Lin, Yi-Ling Lin, Na-Sheng Hsu, Yau-Heiu |
author_sort | Yang, Ming-Hao |
collection | PubMed |
description | We have developed a new binary epitope-presenting CVP platform based on bamboo mosaic virus (BaMV) by using the sortase A (SrtA)-mediated ligation technology. The reconstructed BaMV genome harbors two modifications: 1) a coat protein (CP) with N-terminal extension of the tobacco etch virus (TEV) protease recognition site plus 4 extra glycine (G) residues as the SrtA acceptor; and 2) a TEV protease coding region replacing that of the triple-gene-block proteins. Inoculation of such construct, pKB5G, on Nicotiana benthamiana resulted in the efficient production of filamentous CVPs ready for SrtA-mediated ligation with desired proteins. The second part of the binary platform includes an expression vector for the bacterial production of donor proteins. We demonstrated the applicability of the platform by using the recombinant envelope protein domain III (rEDIII) of Japanese encephalitis virus (JEV) as the antigen. Up to 40% of the BaMV CP subunits in each CVP were loaded with rEDIII proteins in 1 min. The rEDIII-presenting BaMV CVPs (BJLPET5G) could be purified using affinity chromatography. Immunization assays confirmed that BJLPET5G could induce the production of neutralizing antibodies against JEV infections. The binary platform could be adapted as a useful alternative for the development and mass production of vaccine candidates. |
format | Online Article Text |
id | pubmed-8551676 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-85516762021-10-29 Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic Coupling Yang, Ming-Hao Hu, Chung-Chi Wong, Chi-Hzeng Liang, Jian-Jong Ko, Hui-Ying He, Meng-Hsun Lin, Yi-Ling Lin, Na-Sheng Hsu, Yau-Heiu Front Immunol Immunology We have developed a new binary epitope-presenting CVP platform based on bamboo mosaic virus (BaMV) by using the sortase A (SrtA)-mediated ligation technology. The reconstructed BaMV genome harbors two modifications: 1) a coat protein (CP) with N-terminal extension of the tobacco etch virus (TEV) protease recognition site plus 4 extra glycine (G) residues as the SrtA acceptor; and 2) a TEV protease coding region replacing that of the triple-gene-block proteins. Inoculation of such construct, pKB5G, on Nicotiana benthamiana resulted in the efficient production of filamentous CVPs ready for SrtA-mediated ligation with desired proteins. The second part of the binary platform includes an expression vector for the bacterial production of donor proteins. We demonstrated the applicability of the platform by using the recombinant envelope protein domain III (rEDIII) of Japanese encephalitis virus (JEV) as the antigen. Up to 40% of the BaMV CP subunits in each CVP were loaded with rEDIII proteins in 1 min. The rEDIII-presenting BaMV CVPs (BJLPET5G) could be purified using affinity chromatography. Immunization assays confirmed that BJLPET5G could induce the production of neutralizing antibodies against JEV infections. The binary platform could be adapted as a useful alternative for the development and mass production of vaccine candidates. Frontiers Media S.A. 2021-10-14 /pmc/articles/PMC8551676/ /pubmed/34721406 http://dx.doi.org/10.3389/fimmu.2021.739837 Text en Copyright © 2021 Yang, Hu, Wong, Liang, Ko, He, Lin, Lin and Hsu https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Immunology Yang, Ming-Hao Hu, Chung-Chi Wong, Chi-Hzeng Liang, Jian-Jong Ko, Hui-Ying He, Meng-Hsun Lin, Yi-Ling Lin, Na-Sheng Hsu, Yau-Heiu Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic Coupling |
title | Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic Coupling |
title_full | Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic Coupling |
title_fullStr | Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic Coupling |
title_full_unstemmed | Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic Coupling |
title_short | Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic Coupling |
title_sort | convenient auto-processing vector based on bamboo mosaic virus for presentation of antigens through enzymatic coupling |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8551676/ https://www.ncbi.nlm.nih.gov/pubmed/34721406 http://dx.doi.org/10.3389/fimmu.2021.739837 |
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