scholarly journals Investigation of Kluyveromyces marxianus as a novel host for large-scale production of porcine parvovirus virus-like particles

2020 ◽  
Author(s):  
Deqiang Yang ◽  
Lei Chen ◽  
Jinkun Duan ◽  
Yao Yu ◽  
Jungang Zhou ◽  
...  
Keyword(s):  
Kluyveromyces Marxianus ◽  
Large Scale ◽  
Low Cost ◽  
Fetal Loss ◽  
Exchange Chromatography ◽  
Porcine Parvovirus ◽  
Scale Production ◽  
Igg Antibody ◽  
Total Recovery ◽  
Virus Like Particles

Abstract Background: Porcine Parvovirus (PPV) is a Parvovirinae virus that can cause embryonic and fetal loss and death and mummification in affected fetal pigs. Unlike conventional vaccines, virus-like particles (VLPs) inherit the natural structure of their authentic virions and highly immunostimulatory that can induce strong humoral immune and T cell responses with no risk of pathogenicity due to incomplete attenuation or inactivation. Production PPV VLPs is still a challenge using the traditional expression platforms due to the low yield and culture cost. Kluyveromyces marxianus is a safe and fast-growing eukaryote that can yield high biomass in low-cost cultures. In this study, we investigated the production and downstream processes of PPV VLPs in K. marxianus, and the potential for effective stand-alone vaccines.Results: After optimization according to the codon bias of K.marxianus, the VP2 protein from Kresse strain was highly expressed. In 5L fermentator, the yield of PPV VLPs reached 2.5 g/L, qualified by HPLC, using a defined mineral medium after 48h fermentation. Two strategies, cation exchange chromatography coupled with Sephacryl® S-500 HR chromatography for the supernatants of pH adjusted cell lysates and anion exchange chromatography followed by cross-flow diafiltration for the precipitate respectively, were established to purify the intracellular PPV VLPs. The purity PPV VLPs reached about 95%, and total recovery was more than 60%. Sera from the PPV VLPs immunized mice showed high titers of IgG antibody and hemagglutination inhibitions on both swine and guinea pig erythrocytes. Spleen lymphocyte proliferation and cytokines detection suggested the PPV VLPs produced by K.marxianus can provoke the cellular immune and humoral immunity responses.Conclusion: This is the highest production of recombinant PPV VLPs achieved to date. The superiorities, safe, high production, short lead time, and low cost, make K. marxianus a greatly competitive platform in bioproduction of PPV VLPs vaccine.

scholarly journals Investigation of Kluyveromyces marxianus as a novel host for large-scale production of porcine parvovirus virus-like particles

2020 ◽  
Author(s):  
Deqiang Yang ◽  
Lei Chen ◽  
Jinkun Duan ◽  
Yao Yu ◽  
Jungang zhou ◽  
...  
Keyword(s):  
Kluyveromyces Marxianus ◽  
Large Scale ◽  
Low Cost ◽  
Fetal Loss ◽  
Exchange Chromatography ◽  
Porcine Parvovirus ◽  
Scale Production ◽  
Total Recovery ◽  
Virus Like Particles ◽  
Cell Lysates

Abstract Background: Porcine Parvovirus (PPV) is a Parvovirinae virus that can cause embryonic and fetal loss and death and mummification in affected fetal pigs. Unlike conventional vaccines, virus-like particles (VLPs) inherit the natural structure of their authentic virions and highly immunostimulatory that can induce strong humoral immune and T cell responses with no risk of pathogenicity. The production of PPV VLPs is still a challenge based on traditional expression platforms due to their low yields and high culture costs. Kluyveromyces marxianus is a safe and fast-growing eukaryote that can get high biomass with low-cost cultures. In this study, we investigated the expression and downstream processes of PPV VLPs in K. marxianus, and the potential for effective stand-alone vaccines.Results: After optimization according to the codon bias of K.marxianus, the VP2 protein from Kresse strain was highly expressed. In a 5 L fermentator, the yield of PPV VLPs reached 2.5 g/L, quantified by HPLC, using a defined mineral medium after 48 h fermentation. Two strategies were established to purify intracellular PPV VLPs: i) Using the cation exchange chromatography coupled with Sephacryl® S-500 HR chromatography to purify VLPs from the supernatants of pH adjusted cell lysates. ii) Using anion exchange chromatography followed by cross-flow diafiltration to recover the VLPs precipitated in pH adjusted cell lysates. The purity of PPV VLPs reached about 95%, and total recovery was more than 60%. Vaccination of mice with the purified PPV VLPs induced high titers of specific IgG antibodies in sera, and showed hemagglutination inhibitions on both swine and guinea pig erythrocytes. Spleen lymphocyte proliferation and cytokines detection suggested the PPV VLPs produced by K.marxianus provoked the cellular immune and humoral immunity responses in mice.Conclusion: This is the highest production of recombinant PPV VLPs achieved to date. The superiorities, Generally Recognized As Safe (GRAS), high production, short lead time, and low cost, make K. marxianus a greatly competitive platform for bioproduction of PPV VLPs vaccine.

scholarly journals Investigation of Kluyveromyces marxianus as a novel host for large‐scale production of porcine parvovirus virus‐like particles

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Deqiang Yang ◽  
Lei Chen ◽  
Jinkun Duan ◽  
Yao Yu ◽  
Jungang Zhou ◽  
...  
Keyword(s):  
Kluyveromyces Marxianus ◽  
Large Scale ◽  
Low Cost ◽  
Fetal Loss ◽  
Exchange Chromatography ◽  
Porcine Parvovirus ◽  
Scale Production ◽  
Total Recovery ◽  
Virus Like Particles ◽  
Cell Lysates

Abstract Background Porcine Parvovirus (PPV) is a Parvovirinae virus that can cause embryonic and fetal loss and death and mummification in affected fetal pigs. Unlike conventional vaccines, virus-like particles (VLPs) inherit the natural structure of their authentic virions and highly immunostimulatory that can induce strong humoral immune and T cell responses with no risk of pathogenicity. The production of PPV VLPs is still a challenge based on traditional expression platforms due to their low yields and high culture costs. Kluyveromyces marxianus is a safe and fast-growing eukaryote that can get high biomass with low-cost cultures. In this study, we investigated the expression and downstream processes of PPV VLPs in K. marxianus, and the potential for effective stand-alone vaccines. Results After optimization according to the codon bias of K. marxianus, the VP2 protein from Kresse strain was highly expressed. In a 5 L fermentator, the yield of PPV VLPs reached 2.5 g/L, quantified by HPLC, using a defined mineral medium after 48 h fermentation. Two strategies were established to purify intracellular PPV VLPs: (i) Using the cation exchange chromatography coupled with Sephacryl® S-500 HR chromatography to purify VLPs from the supernatants of pH adjusted cell lysates. (ii) Using anion exchange chromatography followed by cross-flow diafiltration to recover the VLPs precipitated in pH adjusted cell lysates. The purity of PPV VLPs reached about 95%, and total recovery was more than 60%. Vaccination of mice with the purified PPV VLPs induced high titers of specific IgG antibodies in sera, and showed hemagglutination inhibitions on both swine and guinea pig erythrocytes. Spleen lymphocyte proliferation and cytokines detection suggested the PPV VLPs produced by K. marxianus provoked the cellular immune and humoral immunity responses in mice. Conclusions This is the highest production of recombinant PPV VLPs achieved to date. The superiorities, Generally Recognized As Safe (GRAS), high production, short lead time, and low cost, make K. marxianus a greatly competitive platform for bioproduction of PPV VLPs vaccine.

scholarly journals Effects of Ag contents on the microstructure and SERS performance of self-grown Ag nanoparticles/Mo–Ag alloy films

2020 ◽  
Vol 9 (1) ◽  
pp. 751-759 ◽  
Author(s):  
Xinxin Lian ◽  
Yuanjiang Lv ◽  
Haoliang Sun ◽  
David Hui ◽  
Guangxin Wang
Keyword(s):  
Ag Nanoparticles ◽  
Large Scale ◽  
Low Cost ◽  
Film Surface ◽  
Time Domain Analysis ◽  
Alloy Film ◽  
Scale Production ◽  
Alloy Films ◽  
Surface Enhanced Raman ◽  
Ag Film

AbstractAg nanoparticles/Mo–Ag alloy films with different Ag contents were prepared on polyimide by magnetron sputtering. The effects of Ag contents on the microstructure of self-grown Ag nanoparticles/Mo–Ag alloy films were investigated using XRD, FESEM, EDS and TEM. The Ag content plays an important role in the size and number of uniformly distributed Ag nanoparticles spontaneously formed on the Mo–Ag alloy film surface, and the morphology of the self-grown Ag nanoparticles has changed significantly. Additionally, it is worth noting that the Ag nanoparticles/Mo–Ag alloy films covered by a thin Ag film exhibits highly sensitive surface-enhanced Raman scattering (SERS) performance. The electric field distributions were calculated using finite-difference time-domain analysis to further prove that the SERS enhancement of the films is mainly determined by “hot spots” in the interparticle gap between Ag nanoparticles. The detection limit of the Ag film/Ag nanoparticles/Mo–Ag alloy film for Rhodamine 6G probe molecules was 5 × 10−14 mol/L. Therefore, the novel type of the Ag film/Ag nanoparticles/Mo–Ag alloy film can be used as an ideal SERS-active substrate for low-cost and large-scale production.

scholarly journals Thermoelectric Performance of Mechanically Mixed BixSb2-xTe3—ABS Composites

Materials ◽  
2021 ◽  
Vol 14 (7) ◽  
pp. 1706
Author(s):  
Zacharias Viskadourakis ◽  
Argiri Drymiskianaki ◽  
Vassilis M. Papadakis ◽  
Ioanna Ioannou ◽  
Theodora Kyratsi ◽  
...  
Keyword(s):  
Polymer Composites ◽  
Large Scale ◽  
Low Cost ◽  
Acrylonitrile Butadiene Styrene ◽  
Thermoelectric Performance ◽  
Scale Production ◽  
Bismuth Antimony Telluride ◽  
Mechanical Mixing ◽  
Large Scale Production ◽  
Bi Doping

In the current study, polymer-based composites, consisting of Acrylonitrile Butadiene Styrene (ABS) and Bismuth Antimony Telluride (BixSb2−xTe3), were produced using mechanical mixing and hot pressing. These composites were investigated regarding their electrical resistivity and Seebeck coefficient, with respect to Bi doping and BixSb2-xTe3 loading into the composite. Experimental results showed that their thermoelectric performance is comparable—or even superior, in some cases—to reported thermoelectric polymer composites that have been produced using other complex techniques. Consequently, mechanically mixed polymer-based thermoelectric materials could be an efficient method for low-cost and large-scale production of polymer composites for potential thermoelectric applications.

scholarly journals Structural Insight into CVB3-VLP Non-Adjuvanted Vaccine

2020 ◽  
Vol 8 (9) ◽  
pp. 1287
Author(s):  
Minna M. Hankaniemi ◽  
Mo A. Baikoghli ◽  
Virginia M. Stone ◽  
Li Xing ◽  
Outi Väätäinen ◽  
...  
Keyword(s):  
Exchange Chromatography ◽  
Scale Production ◽  
Igg Antibody ◽  
Purification Method ◽  
Microscopy Imaging ◽  
Single Particle Reconstruction ◽  
Enhanced Production ◽  
Chronic Myocarditis ◽  
Flow Filtration ◽  
Antibody Levels

Coxsackievirus B (CVB) enteroviruses are common pathogens that can cause acute and chronic myocarditis, dilated cardiomyopathy, aseptic meningitis, and they are hypothesized to be a causal factor in type 1 diabetes. The licensed enterovirus vaccines and those currently in clinical development are traditional inactivated or live attenuated vaccines. Even though these vaccines work well in the prevention of enterovirus diseases, new vaccine technologies, like virus-like particles (VLPs), can offer important advantages in the manufacturing and epitope engineering. We have previously produced VLPs for CVB3 and CVB1 in insect cells. Here, we describe the production of CVB3-VLPs with enhanced production yield and purity using an improved purification method consisting of tangential flow filtration and ion exchange chromatography, which is compatible with industrial scale production. We also resolved the CVB3-VLP structure by Cryo-Electron Microscopy imaging and single particle reconstruction. The VLP diameter is 30.9 nm on average, and it is similar to Coxsackievirus A VLPs and the expanded enterovirus cell-entry intermediate (the 135s particle), which is ~2 nm larger than the mature virion. High neutralizing and total IgG antibody levels, the latter being a predominantly Th2 type (IgG1) phenotype, were detected in C57BL/6J mice immunized with non-adjuvanted CVB3-VLP vaccine. The structural and immunogenic data presented here indicate the potential of this improved methodology to produce highly immunogenic enterovirus VLP-vaccines in the future.

scholarly journals Extensive Screening of Green Solvents for Safe and Sustainable UiO-66 Synthesis

2020 ◽  
Author(s):  
Diletta Morelli Venturi ◽  
Filippo Campana ◽  
Fabio Marmottini ◽  
Ferdinando Costantino ◽  
Luigi Vaccaro
Keyword(s):  
Large Scale ◽  
Low Cost ◽  
Metal Organic Framework ◽  
Dimethyl Formamide ◽  
Scale Production ◽  
Green Solvents ◽  
High Quality ◽  
Color Code ◽  
Large Scale Production ◽  
Metal Organic

<p>Zirconium based Metal-Organic Framework UiO-66 is to date considered one of the benchmark compound among stable MOFs and it has attracted a huge attention for its employment in many strategic applications. Large scale production of UiO-66 for industrial purposes requires the use of safe and green solvents, fulfilling the green chemistry principles and able to replace the use of <i>N,N</i>-Dimethyl-Formamide (DMF), which, despite its toxicity, is still considered the most efficient solvent for obtaining UiO-66 of high quality. Herein we report on a survey of about 40 different solvents with different polarity, boiling point and acidity, used for the laboratory scale synthesis of high quality UiO-66 crystals. The solvents were chosen according the European REACH Regulation 1907/2006 among those having low cost, low toxicity and fully biodegradable. Concerning MOF synthesis, the relevant parameters chosen for establishing the quality of the results obtained are the degree are the crystallinity, microporosity and specific surface area, yield and solvent recyclability. Taking into account also the chemical physical properties of all the solvents, a color code was assigned in order to give a final green assessment for the UiO-66 synthesis. Defectivity of the obtained products, the use of acidic modulators and the use of alternative Zr-salts have been also taken into consideration. Preliminary results lead to conclude that GVL (γ-valerolactone) is among the most promising solvents for replacing DMF in UiO-66 MOF synthesis. </p>

scholarly journals Low-cost Large Scale Vermicompost Unit

2021 ◽  
Vol 2115 (1) ◽  
pp. 012026
Author(s):  
Sonam Solanki ◽  
Gunendra Mahore
Keyword(s):  
Large Scale ◽  
Low Cost ◽  
Small Scale ◽  
Scale Production ◽  
Key Innovation ◽  
Main Challenge ◽  
Long Run ◽  
Large Scale Production ◽  
Term Maintenance

Abstract In the current process of producing vermicompost on a large-scale, the main challenge is to keep the worms alive. This is achieved by maintaining temperature and moisture in their living medium. It is a difficult task to maintain these parameters throughout the process. Currently, this is achieved by building infrastructure but this method requires a large initial investment and long-run maintenance. Also, these methods are limited to small-scale production. For large-scale production, a unit is developed which utilises natural airflow with water and automation. The main aim of this unit is to provide favourable conditions to worms in large-scale production with very low investment and minimum maintenance in long term. The key innovation of this research is that the technology used in the unit should be practical and easy to adopt by small farmers. For long-term maintenance of the technology lesser number of parts are used.

scholarly journals Extensive Screening of Green Solvents for Safe and Sustainable UiO-66 Synthesis

2020 ◽  
Author(s):  
Diletta Morelli Venturi ◽  
Filippo Campana ◽  
Fabio Marmottini ◽  
Ferdinando Costantino ◽  
Luigi Vaccaro
Keyword(s):  
Large Scale ◽  
Low Cost ◽  
Metal Organic Framework ◽  
Dimethyl Formamide ◽  
Scale Production ◽  
Green Solvents ◽  
High Quality ◽  
Color Code ◽  
Large Scale Production ◽  
Metal Organic

<p>Zirconium based Metal-Organic Framework UiO-66 is to date considered one of the benchmark compound among stable MOFs and it has attracted a huge attention for its employment in many strategic applications. Large scale production of UiO-66 for industrial purposes requires the use of safe and green solvents, fulfilling the green chemistry principles and able to replace the use of <i>N,N</i>-Dimethyl-Formamide (DMF), which, despite its toxicity, is still considered the most efficient solvent for obtaining UiO-66 of high quality. Herein we report on a survey of about 40 different solvents with different polarity, boiling point and acidity, used for the laboratory scale synthesis of high quality UiO-66 crystals. The solvents were chosen according the European REACH Regulation 1907/2006 among those having low cost, low toxicity and fully biodegradable. Concerning MOF synthesis, the relevant parameters chosen for establishing the quality of the results obtained are the degree are the crystallinity, microporosity and specific surface area, yield and solvent recyclability. Taking into account also the chemical physical properties of all the solvents, a color code was assigned in order to give a final green assessment for the UiO-66 synthesis. Defectivity of the obtained products, the use of acidic modulators and the use of alternative Zr-salts have been also taken into consideration. Preliminary results lead to conclude that GVL (γ-valerolactone) is among the most promising solvents for replacing DMF in UiO-66 MOF synthesis. </p>

A Ceramic Tubular Probe For Online Substance Concentration Measurement, Manufactured By Ceramic Injection Molding

2012 ◽  
Vol 2012 (1) ◽  
pp. 000604-000608
Author(s):  
Matthias Hartmann ◽  
Bertram Schmidt
Keyword(s):  
Injection Molding ◽  
Zirconium Oxide ◽  
Large Scale ◽  
Low Cost ◽  
Tetragonal Zirconia ◽  
Scale Production ◽  
Ceramic Injection Molding ◽  
Substance Concentration ◽  
Sensor Effect ◽  
Concentration Measurements

The current research presents recent respective to the work development of a ceramic tubular probe for online substance concentration measurements. The aim was to develop a robust and acid-resistant sensor device, which can be easily included in existing procedural pipeline systems. To archive those goals a lot of factors had to be checked. For the substance concentration measurements a capacitive sensor effect was chosen. With this method even low substance concentrations down to one-tenth of a per cent can be indentified. For the package material zirconium oxide (tetragonal zirconia polycrystal – TZP) was used. Zirconium oxide is a technical ceramic which is wear-resistant, acid-resistant, has a low thermal conductivity, is electrically isolating and can be uses in a ceramic injection molding (CIM) process. In the phase of the sensor design process multiple geometries for the sensor effect and integration space for the evaluation electronics had to be considered. A standardized DN 10 DIN 32676 flanged joint was also added for an unproblematic connection to the pipelines. All these needed geometries had to be integrated into one ceramic element. As a result of these requirements a 3D CAD model of the sensor element was designed. The CAD-file has shown that there was only the CIM technology left to comprehend developed sensor geometry. CIM is a low cost process for large-scale production which is distinguished by high size accuracy. In the CIM process the material shrinkage, this is caused by the needed debindering and sintering steps, had to be considered. The developed ceramic tubular probe was successfully tested in multiple fluidic systems. It has left the test phase and is now ready for maturity phase.

scholarly journals Biomass and lipid induction strategies in microalgae for biofuel production and other applications

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Hossein Alishah Aratboni ◽  
Nahid Rafiei ◽  
Raul Garcia-Granados ◽  
Abbas Alemzadeh ◽  
José Rubén Morones-Ramírez
Keyword(s):  
Lipid Content ◽  
Large Scale ◽  
Fossil Fuels ◽  
Low Cost ◽  
Lipid Production ◽  
Arable Land ◽  
Economic Cost ◽  
Biofuel Production ◽  
Scale Production ◽  
Microalgae Biomass

Abstract The use of fossil fuels has been strongly related to critical problems currently affecting society, such as: global warming, global greenhouse effects and pollution. These problems have affected the homeostasis of living organisms worldwide at an alarming rate. Due to this, it is imperative to look for alternatives to the use of fossil fuels and one of the relevant substitutes are biofuels. There are different types of biofuels (categories and generations) that have been previously explored, but recently, the use of microalgae has been strongly considered for the production of biofuels since they present a series of advantages over other biofuel production sources: (a) they don’t need arable land to grow and therefore do not compete with food crops (like biofuels produced from corn, sugar cane and other plants) and; (b) they exhibit rapid biomass production containing high oil contents, at least 15 to 20 times higher than land based oleaginous crops. Hence, these unicellular photosynthetic microorganisms have received great attention from researches to use them in the large-scale production of biofuels. However, one disadvantage of using microalgae is the high economic cost due to the low-yields of lipid content in the microalgae biomass. Thus, development of different methods to enhance microalgae biomass, as well as lipid content in the microalgae cells, would lead to the development of a sustainable low-cost process to produce biofuels. Within the last 10 years, many studies have reported different methods and strategies to induce lipid production to obtain higher lipid accumulation in the biomass of microalgae cells; however, there is not a comprehensive review in the literature that highlights, compares and discusses these strategies. Here, we review these strategies which include modulating light intensity in cultures, controlling and varying CO2 levels and temperature, inducing nutrient starvation in the culture, the implementation of stress by incorporating heavy metal or inducing a high salinity condition, and the use of metabolic and genetic engineering techniques coupled with nanotechnology.

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