Jafar Vatan dost; Shokofeh Hasanabadi
Volume 23, Issue 1 , May and June 2016, , Pages 169-182
Abstract
Nowadays by genetic engineering, recombinant proteins can be produced massively inresponse to demands of industry.Proteins can be expressed in various expression systems and according to the protein type, the expression system can be cell free system, prokaryotic or eukaryotic-based system. For production ...
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Nowadays by genetic engineering, recombinant proteins can be produced massively inresponse to demands of industry.Proteins can be expressed in various expression systems and according to the protein type, the expression system can be cell free system, prokaryotic or eukaryotic-based system. For production of pharmaceutical proteins in most cases post-translational modifications are necessary and mammalian expression systems are the first choice in this regard. However, due to their problems in high production of recombinant proteins, insect expression systems can be an alternative suitable to achieve high expression of many recombinant and complex proteins. In recent year, S2 cell line from Drosophilawas used as the host for the expression of human and non-human recombinant proteins. This system has the advantages such as high cell density and ability to produce folded recombinant proteins with accurate post-translational modifications, especially gamma carboxylation, provides the potential for application in industrial and commercial area.
Hossein Honari; Mehdi Baranvand; Mohammad Ebrahim Minaee
Volume 21, Issue 6 , January and February 2015, , Pages 1103-1112
Abstract
Background: Shigellosis is an acute intestinal infection from Shiga toxin and Shiga-like toxin, which is caused by Shigella and enterohemorrhagic Escherichia coli (EHEC) and entrotoxinogenic Escherichia coli (ETEC). The disease has a high prevalence rates in the world and is known as a bioterrorist agent. ...
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Background: Shigellosis is an acute intestinal infection from Shiga toxin and Shiga-like toxin, which is caused by Shigella and enterohemorrhagic Escherichia coli (EHEC) and entrotoxinogenic Escherichia coli (ETEC). The disease has a high prevalence rates in the world and is known as a bioterrorist agent. STxB is a part of Shiga toxin and have the property of immunogenicity.
Materials and Methods: In this experimental study, the vector pET28a (+) containing the stxB gene was used and was transformed into E. coli BL21 DE3. The bacteria were grown on antibiotic medium and were confirmed with direct PCR, protein expression and SDS-PAGE gel. The recombinant protein purified by nickel column and was confirmed with SDS-PAGE gel and immunoblotting. The chitosan nanofibers containing STxB protein were synthesized by the electrospinning device. The Intranasal and injectable prescription of STxB protein and nanofibers containing STxB protein were performed in mice for four consecutive times and their Antibody titer were assessed. By ELISA, increased IgG antibody titers were observed in injectable and nasal view mode, which may not capture antigen by nasal epithelial cells in mice.
Results: By ELISA, The increase in IgG antibody titers was observed in injectable and intranasal states but not in naonnasal one, which may be due to the lack of antigen captured by nasal epithelial cells in mice. Immunized mice were able to tolerate five times of the Shiga toxin LD50 of E. coli O157: H7.
Conclusion: The results of this study indicate that with nasal and injection prescribed of STxB protein, the immunized mice can tolerate E. coli O157: H7 Shiga toxin.