Isolation and characterization of Faecalibacterium prausnitzii extracellular vesicles
Introduction: Extracellular vesicles (EVs) contain active biological compounds which play important roles in biological processes. The secretion of EVs is a common phenomenon which occurs in archaea, bacteria and mammalian cells. The secretion of bacterial EVs has been discovered in various species...
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| Format: | Article |
| Language: | English |
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Pasteur Institute of Iran
2017-11-01
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| Series: | Vaccine Research |
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| Online Access: | http://vacres.pasteur.ac.ir/article-1-112-en.html |
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| author | B Jafari RA Khavari Nejad F Vaziri SD Siadat |
| author_facet | B Jafari RA Khavari Nejad F Vaziri SD Siadat |
| author_sort | B Jafari |
| collection | DOAJ |
| description | Introduction: Extracellular vesicles (EVs) contain active biological compounds which play important roles in biological processes. The secretion of EVs is a common phenomenon which occurs in archaea, bacteria and mammalian cells. The secretion of bacterial EVs has been discovered in various species of both Gram-negative and Gram-positive bacteria. Faecalibacterium prausnitzii is one of the commensal bacteria in human intestinal tract which has potentially therapeutic effects by secretion of bioactive compounds. Within the last few years, many investigations have been performed with respect to extracting and obtaining EVs and as a result, there are many methods to isolate and characterize EVs. The aim of this study was to isolate EVs from F. prausnitzii strain A2-165 and to characterize their physico-chemical properties. Methods: EVs were isolated from F. prausnitzii strain A2-165 using ultracentrifugation and filtration. The EVs of bacterium were then characterized by Scanning Electron Microscopy, SDS-PAGE, Bradford assay, NanoDrop and Limulus Amebocyte Lysate (LAL) test. Results: The extracted EVs were confirmed with the shape of vesicles and the sizes ranging from ~ 30 to 250 nm. Total protein concentration of EVs were ~ 3 mg/ml from two methods; Bradford and NanoDrop, respectively. Protein profile pattern of F. prausnitzii-derived EVs ranged from 11 up to 245 kDa. Endotoxin measurement was 2.04 EU/ml. Conclusion: The results of the current study demonstrated that EVs purity and conformation were acceptable. However, further investigations are necessary to elucidate the safety, efficacy, practicality and mechanism of action of this bacterium EVs in clinical practices, especially as vaccine delivery vehicles in the field of vaccine research. |
| first_indexed | 2024-12-20T20:29:18Z |
| format | Article |
| id | doaj.art-a73b276fbfc945aaa2572c77e9106e3c |
| institution | Directory Open Access Journal |
| issn | 2383-2819 2423-4923 |
| language | English |
| last_indexed | 2024-12-20T20:29:18Z |
| publishDate | 2017-11-01 |
| publisher | Pasteur Institute of Iran |
| record_format | Article |
| series | Vaccine Research |
| spelling | doaj.art-a73b276fbfc945aaa2572c77e9106e3c2022-12-21T19:27:24ZengPasteur Institute of IranVaccine Research2383-28192423-49232017-11-01435154Isolation and characterization of Faecalibacterium prausnitzii extracellular vesiclesB Jafari0RA Khavari Nejad1F Vaziri2SD Siadat3 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. Department of Mycobacteriology and Pulmonary Research, Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran Department of Mycobacteriology and Pulmonary Research, Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran Introduction: Extracellular vesicles (EVs) contain active biological compounds which play important roles in biological processes. The secretion of EVs is a common phenomenon which occurs in archaea, bacteria and mammalian cells. The secretion of bacterial EVs has been discovered in various species of both Gram-negative and Gram-positive bacteria. Faecalibacterium prausnitzii is one of the commensal bacteria in human intestinal tract which has potentially therapeutic effects by secretion of bioactive compounds. Within the last few years, many investigations have been performed with respect to extracting and obtaining EVs and as a result, there are many methods to isolate and characterize EVs. The aim of this study was to isolate EVs from F. prausnitzii strain A2-165 and to characterize their physico-chemical properties. Methods: EVs were isolated from F. prausnitzii strain A2-165 using ultracentrifugation and filtration. The EVs of bacterium were then characterized by Scanning Electron Microscopy, SDS-PAGE, Bradford assay, NanoDrop and Limulus Amebocyte Lysate (LAL) test. Results: The extracted EVs were confirmed with the shape of vesicles and the sizes ranging from ~ 30 to 250 nm. Total protein concentration of EVs were ~ 3 mg/ml from two methods; Bradford and NanoDrop, respectively. Protein profile pattern of F. prausnitzii-derived EVs ranged from 11 up to 245 kDa. Endotoxin measurement was 2.04 EU/ml. Conclusion: The results of the current study demonstrated that EVs purity and conformation were acceptable. However, further investigations are necessary to elucidate the safety, efficacy, practicality and mechanism of action of this bacterium EVs in clinical practices, especially as vaccine delivery vehicles in the field of vaccine research.http://vacres.pasteur.ac.ir/article-1-112-en.htmlfaecalibacterium prausnitziiextracellular vesiclesisolationcharacterizationvaccine vehicle. |
| spellingShingle | B Jafari RA Khavari Nejad F Vaziri SD Siadat Isolation and characterization of Faecalibacterium prausnitzii extracellular vesicles Vaccine Research faecalibacterium prausnitzii extracellular vesicles isolation characterization vaccine vehicle. |
| title | Isolation and characterization of Faecalibacterium prausnitzii extracellular vesicles |
| title_full | Isolation and characterization of Faecalibacterium prausnitzii extracellular vesicles |
| title_fullStr | Isolation and characterization of Faecalibacterium prausnitzii extracellular vesicles |
| title_full_unstemmed | Isolation and characterization of Faecalibacterium prausnitzii extracellular vesicles |
| title_short | Isolation and characterization of Faecalibacterium prausnitzii extracellular vesicles |
| title_sort | isolation and characterization of faecalibacterium prausnitzii extracellular vesicles |
| topic | faecalibacterium prausnitzii extracellular vesicles isolation characterization vaccine vehicle. |
| url | http://vacres.pasteur.ac.ir/article-1-112-en.html |
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