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...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2021-10-01
|
| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2021.739837/full |
| _version_ | 1818588005196103680 |
|---|---|
| author | Ming-Hao Yang Chung-Chi Hu Chung-Chi Hu Chi-Hzeng Wong Jian-Jong Liang Hui-Ying Ko Meng-Hsun He Yi-Ling Lin Yi-Ling Lin Na-Sheng Lin Yau-Heiu Hsu Yau-Heiu Hsu |
| author_facet | Ming-Hao Yang Chung-Chi Hu Chung-Chi Hu Chi-Hzeng Wong Jian-Jong Liang Hui-Ying Ko Meng-Hsun He Yi-Ling Lin Yi-Ling Lin Na-Sheng Lin Yau-Heiu Hsu Yau-Heiu Hsu |
| author_sort | Ming-Hao Yang |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-16T09:17:52Z |
| format | Article |
| id | doaj.art-e6b6d86b2d7747a28fc3a0434b02390c |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-16T09:17:52Z |
| publishDate | 2021-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-e6b6d86b2d7747a28fc3a0434b02390c2022-12-21T22:36:51ZengFrontiers Media S.A.Frontiers in Immunology1664-32242021-10-011210.3389/fimmu.2021.739837739837Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic CouplingMing-Hao Yang0Chung-Chi Hu1Chung-Chi Hu2Chi-Hzeng Wong3Jian-Jong Liang4Hui-Ying Ko5Meng-Hsun He6Yi-Ling Lin7Yi-Ling Lin8Na-Sheng Lin9Yau-Heiu Hsu10Yau-Heiu Hsu11Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, TaiwanGraduate Institute of Biotechnology, National Chung Hsing University, Taichung, TaiwanAdvanced Plant Biotechnology Center, National Chung Hsing University, Taichung, TaiwanGraduate Institute of Biotechnology, National Chung Hsing University, Taichung, TaiwanInstitute of Biomedical Sciences, Academia Sinica, Taipei, TaiwanInstitute of Biomedical Sciences, Academia Sinica, Taipei, TaiwanInstitute of Plant and Microbial Biology, Academia Sinica, Taipei, TaiwanInstitute of Biomedical Sciences, Academia Sinica, Taipei, TaiwanBiomedical Translation Research Center, Academia Sinica, Taipei, TaiwanInstitute of Plant and Microbial Biology, Academia Sinica, Taipei, TaiwanGraduate Institute of Biotechnology, National Chung Hsing University, Taichung, TaiwanAdvanced Plant Biotechnology Center, National Chung Hsing University, Taichung, TaiwanWe 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.https://www.frontiersin.org/articles/10.3389/fimmu.2021.739837/fullBamboo mosaic virus (BaMV)chimeric virus particle (CVP)nanoparticlevirus-like particle (VLP)antigen presentation platformTobacco etch virus protease (TEVp) |
| spellingShingle | Ming-Hao Yang Chung-Chi Hu Chung-Chi Hu Chi-Hzeng Wong Jian-Jong Liang Hui-Ying Ko Meng-Hsun He Yi-Ling Lin Yi-Ling Lin Na-Sheng Lin Yau-Heiu Hsu Yau-Heiu Hsu Convenient Auto-Processing Vector Based on Bamboo Mosaic Virus for Presentation of Antigens Through Enzymatic Coupling Frontiers in Immunology Bamboo mosaic virus (BaMV) chimeric virus particle (CVP) nanoparticle virus-like particle (VLP) antigen presentation platform Tobacco etch virus protease (TEVp) |
| 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 | Bamboo mosaic virus (BaMV) chimeric virus particle (CVP) nanoparticle virus-like particle (VLP) antigen presentation platform Tobacco etch virus protease (TEVp) |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2021.739837/full |
| work_keys_str_mv | AT minghaoyang convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT chungchihu convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT chungchihu convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT chihzengwong convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT jianjongliang convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT huiyingko convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT menghsunhe convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT yilinglin convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT yilinglin convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT nashenglin convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT yauheiuhsu convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling AT yauheiuhsu convenientautoprocessingvectorbasedonbamboomosaicvirusforpresentationofantigensthroughenzymaticcoupling |