Preparation of outer membrane vesicles from rabbit Bordetella bronchiseptica and their protein composition analysis
This study aimed to establish the optimal preparation method of outer membrane vesicles (OMVs) from rabbit Bordetella bronchiseptica (Bb). The FX-1 strain of rabbit Bb was used as the research object. Centrifugal ultrafiltration and ultrasonication were compared, and followed by the optimization of...
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| Format: | Article |
| Language: | English |
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Zhejiang University Press
2021-04-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2020.05.190 |
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| author | NAN Li HUANG Ye’e XIAO Chenwen WANG Zhipeng WEI Qiang JI Quan’an LI Ke LIU Yan BAO Guolian |
| author_facet | NAN Li HUANG Ye’e XIAO Chenwen WANG Zhipeng WEI Qiang JI Quan’an LI Ke LIU Yan BAO Guolian |
| author_sort | NAN Li |
| collection | DOAJ |
| description | This study aimed to establish the optimal preparation method of outer membrane vesicles (OMVs) from rabbit Bordetella bronchiseptica (Bb). The FX-1 strain of rabbit Bb was used as the research object. Centrifugal ultrafiltration and ultrasonication were compared, and followed by the optimization of the preparation technics, such as culture time, antibiotic addition and filtering method. The physical and chemical properties of OMVs were examined with transmission electron microscope (TEM), scanning electron microscope (SEM) and nanoparticle tracking analysis (NTA), etc. The protein compositions were analysed with Bradford protein quantification, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results showed that centrifugal ultrafiltration was a better preparation method of OMVs, and the optimum culture time, cefalexin concentration and filtering method were 18 h, 64 μg/mL and filtration though 0.45 µm membrane once, respectively. The OMVs showed a spherical shape with the diameter of 127.83 nm±0.68 nm, containing a range of proteins associated with virulence and infection mechanisms. This preparation technique could significantly promote the production of OMVs, which making industrial production possible. Furthermore, analysis of the protein composition lays a foundation for novel subunit vaccine research and development. |
| format | Article |
| id | doaj-art-2c1db5ed43b548639961ce1129ad9c01 |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2021-04-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-2c1db5ed43b548639961ce1129ad9c012025-08-20T03:32:05ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552021-04-014725126010.3785/j.issn.1008-9209.2020.05.19010089209Preparation of outer membrane vesicles from rabbit Bordetella bronchiseptica and their protein composition analysisNAN LiHUANG Ye’eXIAO ChenwenWANG ZhipengWEI QiangJI Quan’anLI KeLIU YanBAO GuolianThis study aimed to establish the optimal preparation method of outer membrane vesicles (OMVs) from rabbit Bordetella bronchiseptica (Bb). The FX-1 strain of rabbit Bb was used as the research object. Centrifugal ultrafiltration and ultrasonication were compared, and followed by the optimization of the preparation technics, such as culture time, antibiotic addition and filtering method. The physical and chemical properties of OMVs were examined with transmission electron microscope (TEM), scanning electron microscope (SEM) and nanoparticle tracking analysis (NTA), etc. The protein compositions were analysed with Bradford protein quantification, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results showed that centrifugal ultrafiltration was a better preparation method of OMVs, and the optimum culture time, cefalexin concentration and filtering method were 18 h, 64 μg/mL and filtration though 0.45 µm membrane once, respectively. The OMVs showed a spherical shape with the diameter of 127.83 nm±0.68 nm, containing a range of proteins associated with virulence and infection mechanisms. This preparation technique could significantly promote the production of OMVs, which making industrial production possible. Furthermore, analysis of the protein composition lays a foundation for novel subunit vaccine research and development.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2020.05.190bacterial outer membrane vesicles<italic>Bordetella bronchiseptica</italic> preparation technicprotein composition analysis |
| spellingShingle | NAN Li HUANG Ye’e XIAO Chenwen WANG Zhipeng WEI Qiang JI Quan’an LI Ke LIU Yan BAO Guolian Preparation of outer membrane vesicles from rabbit Bordetella bronchiseptica and their protein composition analysis 浙江大学学报. 农业与生命科学版 bacterial outer membrane vesicles <italic>Bordetella bronchiseptica</italic> preparation technic protein composition analysis |
| title | Preparation of outer membrane vesicles from rabbit Bordetella bronchiseptica and their protein composition analysis |
| title_full | Preparation of outer membrane vesicles from rabbit Bordetella bronchiseptica and their protein composition analysis |
| title_fullStr | Preparation of outer membrane vesicles from rabbit Bordetella bronchiseptica and their protein composition analysis |
| title_full_unstemmed | Preparation of outer membrane vesicles from rabbit Bordetella bronchiseptica and their protein composition analysis |
| title_short | Preparation of outer membrane vesicles from rabbit Bordetella bronchiseptica and their protein composition analysis |
| title_sort | preparation of outer membrane vesicles from rabbit bordetella bronchiseptica and their protein composition analysis |
| topic | bacterial outer membrane vesicles <italic>Bordetella bronchiseptica</italic> preparation technic protein composition analysis |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2020.05.190 |
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