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Production of recombinant CAMP – Sialidase protein and preparation of chitosan nanoparticles carrying this protein to be used as a candidate for vaccines targeting Propionibacterium acnes
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P Zargham   , M Heshmati , K Mansouri , J Amani , J Salimian , A Ahmadi * |
| 4. Molecular Biology Research Center, Institute of Biology and Food Poisoning, Baqiyatallah University of Medical Sciences, Tehran, I.R.Iran. , ahmadi1919@gmail.com |
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Abstract: (4164 Views) |
BACKGROUND AND OBJECTIVE: Acne vulgaris is one of the most common skin diseases that imposes too much mental pressure and high costs on patients. The existing treatments have low efficacy and include antibiotics and anti-inflammatory drugs, which have many complications due to the chronic nature of the disease, especially at young age, including antibiotic resistance and allergic reactions. For this reason, one of the new therapeutic approaches is the use of a vaccine with the help of the bacterium or its components. The aim of this study is to produce nanoparticles carrying the recombinant CAMP – Sialidase protein as a new chimeric antigen to be used in acne vaccine.
METHODS: To express the recombinant CAMP – Sialidase protein, E. coli BL21 DE3 was used as the host. Purification of protein was done through combined urea/imidazole method and using a nickel-nitroacetic acid column. The recombinant protein was confirmed using Western Blotting by Anti – Histidine Antibody. Then, the loaded nanoparticles were prepared by recombinant protein using ionic gelation technique and tripolyphosphate. Finally, the size and zeta potential of the nanoparticles were determined by the DLS device.
FINDINGS: The recombinant CAMP – Sialidase protein was confirmed after expression and purification. The size and zeta potential of nanoparticles containing recombinant CAMP – Sialidase protein at a concentration of 0.6 mg / ml were determined to be 80 nm and +27 mV, respectively, using the DLS device. The loading rate of the protein in the nanoparticles was found to be 88%.
CONCLUSION: The results show that the recombinant protein is expressed completely and is successfully encapsulated in the chitosan nanoparticles.
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| Keywords: Acne Vulgaris, Camp – Sialidase Protein, Chitosan Nanoparticle, Vaccine |
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Full-Text [PDF 235 kb]
(1291 Downloads)
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Type of Study: Experimental |
Subject:
Nanobiotechnology
Received: 2017/08/6 | Accepted: 2018/03/3 | Published: 2018/03/17
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