Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 12 1 2020 03 03 37 42 Mohammad Sekhavati Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Ashraf Mohabati Mobarez Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Seyed Davar Siadat Microbiology Research Center (MRC) Pasteur, Institute of Iran, Tehran, Iran Mojtaba Noofeli Department of Human Bacterial Vaccine, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran 2019 07 27 2019 09 22 Background and Objectives: There are many pertussis outbreaks which is mainly due to the reduction in the immunity of acellular pertussis (aP) vaccines. Therefore, there is a crucial necessity to develop a new generation of pertussis vaccine. Preceding researches have shown that Bordetella pertussis outer membrane vesicles (OMVs) have appropriate specifications, making them a suitable vaccine candidate against pertussis. Materials and Methods: The OMVs were separated by a new serial ultra centrifugation technique. Transmission electron microscopy (TEM) examination, SDS-PAGE, Western blotting and ELISA assay were used to characterize the OMVs. Results: TEM studies showed the size of the extracted OMVs at 40-200 nm. The presence of pertussis toxin, filamentous hemagglutinin, and pertactin was verified using Western blot and ELISA assay. Conclusion: The presented technique is a simple and effective way to obtain OMVs from Bordetella pertussis. So it can be utilized as an appropriate procedure in the development of an OMV-based vaccine against pertussis. https://ijm.tums.ac.ir/index.php/ijm/article/view/2267 https://ijm.tums.ac.ir/index.php/ijm/article/download/2267/1220