fa Original article Original article <div style="text-align: justify;"><strong>Introduction:</strong> In spite of high vaccination coverage, whooping cough (pertussis) is still a worldwide health problem. The main reason for pertussis outbreak is waning immunity of safer acellular vaccines which have replaced the more reactogenic cellular vaccines. A new generation of pertussis vaccines that is potent and safe is desperately needed to control the disease. Previous studies have indicated that outer membrane vesicles (OMVs) obtained from <em>Bordetella pertussis </em>have desirable characteristics which make them a good candidate for application as pertussis vaccine. They contain surface immunogens in a native structure, are self-adjuvant and are easily uptaken by the antigen presenting cells. <strong>Methods:</strong> <em>B. pertussis</em> Tohama strain was cultured at 35&deg;C in Stainer-Scholte broth. The OMVs were isolated by a new sequential ultracentrifugation method. The extracted OMVs were characterized by electron microscopy, SDS-PAGE and ELISA assays. <strong>R</strong><strong>esults: </strong>The existence of pertussis toxin, filamentous haemagglutinin and a 69-kDa antigen in <em>B. pertussis</em> OMVs was verified using an ELISA assay. Electron microscopy showed the size of these OMV&rsquo;s at 40-200 nm. The ELISA results indicated that the OMVs extracted using this protocol contain major immunogens. <strong>Conclusion: </strong>We report for the first time a simple protocol for the efficient extraction of <em>B. pertussis</em> OMVs. This protocol can be used in the process of making new generations of <em>B. pertussis</em> vaccines.</div> Bordetella pertussis, outer membrane vesicles, extraction, characterization, vaccine 43 46 http://vacres.pasteur.ac.ir/browse.php?a_code=A-10-1-61&slc_lang=fa&sid=1 M Sekhavati m.sekhavati22@gmail.com 0000-0003-1629-5934 No Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. SD Siadat d.siadat@gmail.com 0000-0002-6892-5603 No Microbiology Research Center (MRC) Pasteur Institute of Iran, Tehran, Iran. M Noofeli 0000-0003-4991-4631 No Human Bacterial Vaccine Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran. A Mohebati Mobarez mmmobarez @modares.ac.ir 0000-0002-1224-6134 Yes Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.