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Increased Expression of Flagellins Recombinant Soluble form by Chemical Chaperon
, M.Sc. Thesis Sharif University of Technology ; Yaghmaei, Soheila (Supervisor) ; Tarahomjoo, Shirin (Supervisor)
Abstract
Flagella is the organelle involved in motility of Salmonella and plays an important role in colonization of Salmonella. Flagellins is the structural subunits of flagellar filament that is known as an adjuvant. Recombinant production of flagellin of S. enteritidis is advantageous due to the removal of pathogenic microorganism from the production process and improved process safety. E.coli is an appropriate host for the expression of recombinant proteins. However, its major obstacle is the formation of inclusion bodies. Inclusion bodies are aggregated inactive proteins and the recovery of proteins in the active form from these aggregates usually has a low yield. Therefore, the objective of our...
Chaperones promote remarkable solubilization of salmonella enterica serovar enteritidis flagellin expressed in escherichia coli
, Article Protein and Peptide Letters ; Volume 27, Issue 3 , 2020 , Pages 210-218 ; Sadeghi, Z ; Tarahomjoo, S ; Yaghmaie, S ; Sharif University of Technology
Bentham Science Publishers
2020
Abstract
Background: Flagellin of Salmonella enterica serovar Enteritidis (SEF) stimulates immune responses to both itself and coapplied antigens. It is therefore used in vaccine development and immunotherapy. Removal of pathogenic S. enterica ser. Enteritidis from SEF production process is advantageous due to the process safety improvement. The protein solubility analysis using SDS-PAGE indicated that 53.49% of SEF expressed in Escherichia coli formed inclusion bodies. However, the protein recovery from inclusion bodies requires a complex process with a low yield. Objective: We thus aim to study possibility of enhancing SEF expression in E. coli in soluble form using chemical and molecular...
Effect of cysteine oxidation in SARS-CoV-2 receptor-binding domain on its interaction with two cell receptors: Insights from atomistic simulations
, Article Journal of Chemical Information and Modeling ; Volume 62, Issue 1 , 2022 , Pages 129-141 ; 15499596 (ISSN) ; Privat Maldonado, A ; Yusupov, M ; Rahnama, S ; Bogaerts, A ; Ejtehadi, M. R ; Sharif University of Technology
American Chemical Society
2022
Abstract
Binding of the SARS-CoV-2 S-glycoprotein to cell receptors is vital for the entry of the virus into cells and subsequent infection. ACE2 is the main cell receptor for SARS-CoV-2, which can attach to the C-terminal receptor-binding domain (RBD) of the SARS-CoV-2 S-glycoprotein. The GRP78 receptor plays an anchoring role, which attaches to the RBD and increases the chance of other RBDs binding to ACE2. Although high levels of reactive oxygen and nitrogen species (RONS) are produced during viral infections, it is not clear how they affect the RBD structure and its binding to ACE2 and GRP78. In this research, we apply molecular dynamics simulations to study the effect of oxidation of the highly...
Modeling, simulation, and employing dilution–dialysis microfluidic chip (DDMC) for heightening proteins refolding efficiency
, Article Bioprocess and Biosystems Engineering ; Volume 41, Issue 5 , 2018 , Pages 707-714 ; 16157591 (ISSN) ; Masoudi, M. M ; Shamloo, A ; Habibi Rezaei, M ; Moosavi Movahedi, A. A ; Sharif University of Technology
Springer Verlag
2018
Abstract
Miniaturized systems based on the principles of microfluidics are widely used in various fields, such as biochemical and biomedical applications. Systematic design processes are demanded the proper use of these microfluidic devices based on mathematical simulations. Aggregated proteins (e.g., inclusion bodies) in solution with chaotropic agents (such as urea) at high concentration in combination with reducing agents are denatured. Refolding methods to achieve the native proteins from inclusion bodies of recombinant protein relying on denaturant dilution or dialysis approaches for suppressing protein aggregation is very important in the industrial field. In this paper, a modeling approach is...