Designing a chimeric subunit vaccine for influenza virus, based on HA2, M2e and CTxB: a bioinformatics study

Background: Type A influenza viruses are contagious and even life-threatening if left untreated. So far, no broadly protective vaccine is available due to rapid antigenic changes and emergence of new subtypes of influenza virus. In this study, we exploited bioinformatics tools in order to design a subunit chimeric vaccine from the antigenic and highly conserved regions of HA and M2 proteins of H7N9 subtype of influenza virus. We used mucosal adjuvant candidates, including CTxB, STxB, ASP-1, and LTB to stimulate mucosal immunity and analyzed the combination of HA2, M2e, and the adjuvant. Furthermore, to improve the antigen function and to maintain their three-dimensional structure, 12 different linkers including six rigid linkers and six flexible linkers were used. The 3D structure model was generated using a combination of homology and ab initio modeling methods and the molecular dynamics of the model were analyzed, either. Results: Analysis of different adjuvants showed that using CtxB as an adjuvant, results in higher overall vaccine stability and higher half-life among four adjuvant candidates. Fusion of antigens and the CTxB in the form of M2e-linker-CTxB-linker-HA2 has the most stability and half life compared to other combination forms. Furthermore, the KPKPKP rigid linker showed the best result for this candidate vaccine among 12 analyzed linkers. The changes in the vaccine 3D structure made by linker insertion found to be negligible, however, although small, the linker insertion between the antigens causes the structure to change slightly. Eventually, using predictive tools such as Ellipro, NetMHCpan I and II, CD4episcore, CTLpred, BepiPred and other epitope analyzing tools, we analyzed the conformational and linear epitopes of the vaccine. The solubility, proteasome cleavage sites, peptidase and potential chemical cutters, codon optimization, post translational modification were also carried out on the final vaccine. Conclusions: It is concluded that M2e-Linker-CTxB-Linker-HA2 combination of chimeric vaccine retains its 3D structure and antigenicity when KPKPKP used as linker and CTxB used as adjuvant. © 2020, The Author(s)

Designer Exosomes: A New Platform for Biotechnology Therapeutics

Abstract: Desirable features of exosomes have made them a suitable manipulative platform for biomedical applications, including targeted drug delivery, gene therapy, cancer diagnosis and therapy, development of vaccines, and tissue regeneration. Although natural exosomes have various potentials, their clinical application is associated with some inherent limitations. Recently, these limitations inspired various attempts to engineer exosomes and develop designer exosomes. Mostly, designer exosomes are being developed to overcome the natural limitations of exosomes for targeted delivery of drugs and functional molecules to wounds, neurons, and the cardiovascular system for healing of damage. In this review, we summarize the possible improvements of natural exosomes by means of two main approaches: parental cell-based or pre-isolation exosome engineering and direct or post-isolation exosome engineering. Parental cell-based engineering methods use genetic engineering for loading of therapeutic molecules into the lumen or displaying them on the surface of exosomes. On the other hand, the post-isolation exosome engineering approach uses several chemical and mechanical methods including click chemistry, cloaking, bio-conjugation, sonication, extrusion, and electroporation. This review focuses on the latest research, mostly aimed at the development of designer exosomes using parental cell-based engineering and their application in cancer treatment and regenerative medicine. Graphic Abstract: Figure not available: see fulltext. © 2020, Springer Nature Switzerland AG

An investigation into the roles of chlorides and sulphate salts on the performance of low salinity injection in sandstone reservoirs : experimental approach

Numerous studies have been carried out to ascertain the mechanisms of low salinity and smart water flooding technique for improved oil recovery. Focus were often on brine composition and, specifically the cationic content in sandstone reservoirs. Given the importance of the salt composition and concentration, tweaking the active ions which are responsible for the fluids-rock equilibrium will bring into effect numerous mechanisms of displacement which have been extensively debated. This experimental study, however, was carried out to evaluate the extent of the roles of chloride and sulphate-based brines in improved oil recovery. To carry this out, 70,000 ppm sulphates and chloride-based brines were prepared to simulate formation water and 5,000ppm brines of the same species as low salinity displacement fluids. Core flooding process was used to simulate the displacement of oil by using four (4) native sandstones core samples, obtained from Burgan oil field in Kuwait, at operating conditions of 1500 psig and 50oC. The core samples were injected with 70,000 ppm chloride and sulphates and subsequently flooded with the 5,000 ppm counterparts in a forced imbibition process. Separate evaluations of chloride and sulphate-based brines were carried out to investigate the displacement efficiencies of each brine species. The results showed that the in both high and low salinity displacement tests, the SO4 brine presented better recovery of up to 89% of the initial oil saturation (Soi). Several mechanisms of displacement were observed to be responsible for improved recovery during SO4 brine displacement. IFT measurement experiments also confirmed that there was reduction in IFT at test conditions between SO4 brine and oil and visual inspection of the effluent showed a degree emulsification of oil and brines. Changes in pH were observed in the low salinity flooding and negligible changes were noticed in the high salinity floods. These results provide an insight into the roles of chloride and sulphate ions in the design of smart “designer” water and low salinity injection scenarios

Morphology and electrochemical properties of a gel blend polymer electrolyte based on PVDF-HFP/PEO blend

Hypothesis: In recent years, gel polymer electrolytes (quasi-solid state electrolytes) have attracted great attention as a suitable substitute for liquid electrolytes. On the other hand, ionic liquids could dramatically enhance the ionic conductivity of electrolytes. In this work, gel polymer electrolytes based on poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP)/poly(ethylene oxide) (PEO) blends (for application in dye-sensitized solar cells (DSSCs)) and imidazoliumbased ionic liquids were prepared. It is supposed that blending these two polymers could reduce the degree of crystallization and increase the porosity of the electrolyte blend to yield a higher electrolyte uptake and ionic conductivity. Methods: Polymer blend electrolytes were prepared in different blend ratios and in the presence of either one of the ionic liquids including BMII or BMIMBF4 through phase inversion method and their properties were investigated by differential scanning calorimetry (DSC), mercury porosimetry, electrolyte uptake, and morphology to optimize the blend ratio. Findings: It was found that the blend ratio of 60/40 (w/w) PVDF-HFP/PEO has the highest porosity and electrolyte uptake. Crystallization investigations by DSC showed that there is a direct relationship between the decrease of crystallinity of two polymers and the increment of electrolyte ionic conductivity. Electrolyte uptake gradually increased with increasing PEO component concentration up to 40 wt%, and reached a maximum of 98.49% and 89.48% for BMIMBF4 and BMII, respectively. Beyond this concentration, a decrease in electrolyte uptake was seen, which is an undesirable feature for the produced samples. In this blend ratio ionic conductivity was measured as 2.07 mS/cm and 1.78 mS/cm for PVDF-HFP/PEO/BMIMBF4 and PVDF-HFP/ PEO/BMII electrolytes, respectively

Opportunities and challenges of the tag-assisted protein purification techniques: Applications in the pharmaceutical industry

Tag-assisted protein purification is a method of choice for both academic researches and large-scale industrial demands. Application of the purification tags in the protein production process can help to save time and cost, but the design and application of tagged fusion proteins are challenging. An appropriate tagging strategy must provide sufficient expression yield and high purity for the final protein products while preserving their native structure and function. Thanks to the recent advances in the bioinformatics and emergence of high-throughput techniques (e.g. SEREX), many new tags are introduced to the market. A variety of interfering and non-interfering tags have currently broadened their application scope beyond the traditional use as a simple purification tool. They can take part in many biochemical and analytical features and act as solubility and protein expression enhancers, probe tracker for online visualization, detectors of post-translational modifications, and carrier-driven tags. Given the variability and growing number of the purification tags, here we reviewed the protein- and peptide-structured purification tags used in the affinity, ion-exchange, reverse phase, and immobilized metal ion affinity chromatographies. We highlighted the demand for purification tags in the pharmaceutical industry and discussed the impact of self-cleavable tags, aggregating tags, and nanotechnology on both the column-based and column-free purification techniques. © 202