Volume 22, Issue 2 (2019)
mjms 2019, 22(2): 69-75 |
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Hosseini Z, Amani J, Hosseini F. Expression of the Recombinant Protein Containing CfaB, ST, CfaE, and LtB from Enterotoxigenic Escherichia coli and Loading It in Chitosan Nanoparticles. mjms 2019; 22 (2) :69-75
URL: http://mjms.modares.ac.ir/article-30-22664-en.html
URL: http://mjms.modares.ac.ir/article-30-22664-en.html
1- Cell & Molecular Biology-Biotechnology Department, Biological Sciences Faculty, Tehran North Branch, Islamic Azad University, Tehran, Iran
2- Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran , jafar.amani@gmail.com
2- Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran , jafar.amani@gmail.com
Abstract: (7095 Views)
Aims: Enterotoxigenic Escherichia coli (ETEC) is the most important bacteria causing traveler’s diarrhea. The bacterium has several virulence factors, including colonization factors (CFs) or Escherichia coli adhesins, heat-labile (LT), and heat-stable (ST) toxins. The design and production of vaccine against this disease is one of the goals of the World Health Organization due to increased antibiotic resistance and a reduction of healthy water sources. An effective subunit vaccine against ETEC could include a toxoid from both toxins and colonization factors. The aim of the current study was to express, purify, and encapsulate the recombinant protein in chitosan nanoparticles.
Materials and Methods: In the present experimental study, the E. coli BL21DE3 harbring pET-28a-cscl vector was used. The chimeric cscl gene is composed of cfab along with st toxin, cfae, and ltb. After the expression and purification of recombinant protein, using Ni-NTA column, Western blotting was performed with anti-His antibody. Then, the CSCl protein was encapsulated in chitosan nanoparticles and the particle size was measured.
Findings: The recombinant CSCL protein was purified by Ni-NTA column and urea denaturation method. Then, this purified protein (~57kDa) was confirmed by Western blotting and the size of the nanoparticles was estimated as 112.0 nm with 98.8% of encapsulation efficiency.
Conclusion: With some advantages, including the presence of surface and important antigens of ETEC and encapsulating in chitosan nanoparticles, the CSCL recombinant protein can be considered as a candidate for producing oral nanovaccine and stimulating of mucosal and systemic immune response.
Materials and Methods: In the present experimental study, the E. coli BL21DE3 harbring pET-28a-cscl vector was used. The chimeric cscl gene is composed of cfab along with st toxin, cfae, and ltb. After the expression and purification of recombinant protein, using Ni-NTA column, Western blotting was performed with anti-His antibody. Then, the CSCl protein was encapsulated in chitosan nanoparticles and the particle size was measured.
Findings: The recombinant CSCL protein was purified by Ni-NTA column and urea denaturation method. Then, this purified protein (~57kDa) was confirmed by Western blotting and the size of the nanoparticles was estimated as 112.0 nm with 98.8% of encapsulation efficiency.
Conclusion: With some advantages, including the presence of surface and important antigens of ETEC and encapsulating in chitosan nanoparticles, the CSCL recombinant protein can be considered as a candidate for producing oral nanovaccine and stimulating of mucosal and systemic immune response.
Keywords: Enterotoxigenic Escherichia coli, Heat-labile toxin, Heat-stable toxin, Chitosan nanoparticles, Escherichia coli adhesins, Traveler’s diarrhea
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