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...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2021-04-01
|
| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2020.05.190 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Summary: | 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. |
|---|---|
| ISSN: | 1008-9209 2097-5155 |