scholarly journals Lessons Learned in Protein Precipitation Using a Membrane Emulsification Technique to Produce Reversible and Uniform Microbeads

Pharmaceutics ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1738
Author(s):  
Sang-Koo Park ◽  
Ga Yeon Noh ◽  
Hyun Woo Yu ◽  
Eun Chae Lee ◽  
Junoh Jeong ◽  
...  
Keyword(s):  
Intravenous Immunoglobulin ◽  
Porous Glass ◽  
Manufacturing Process ◽  
Protein Concentration ◽  
Lessons Learned ◽  
The Other ◽  
Silicone Resin ◽  
Hydrophobic Modification ◽  
Hydrophobically Modified ◽  
Model Protein

The effects of the manufacturing process and the regeneration of Shirasu porous glass (SPG) membranes were investigated on the reproducibility of protein precipitants, termed protein microbeads. Intravenous immunoglobulin (IVIG) was selected as a model protein to produce its microbeads in seven different cases. The results showed that the hydrophobically modified SPG membrane produced finer microbeads than the hydrophilic SPG membrane, but this was inconsistent when using the general regeneration method. Its reproducibility was determined to be mostly dependent on rinsing the SPG membrane prior to the modification and on the protein concentration used for emulsification. The higher concentration could foul and plug the membrane during protein release and thus the membrane must be washed thoroughly before hydrophobic modification. Moreover, the membrane regenerated by silicone resin dissolved in ethanol had better reproducibility than silicone resin dissolved in water. On the other hand, rinsing the protein precipitant with cold ethanol after the emulsification was not favorable and induced protein aggregation. With the addition of trehalose, the purity of the IVIG microbeads was almost the same as before microbeadification. Therefore, the regeneration method, protein concentration, and its stabilizer are key to the success of protein emulsification and precipitation using the SPG membrane.

scholarly journals Anti-EGFR VHH Antibody under Thermal Stress is Better Solubilized with a Lysine than with an Arginine SEP Tag

Biomolecules ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 810
Author(s):  
Md. Golam Kibria ◽  
Akari Fukutani ◽  
Yoko Akazawa-Ogawa ◽  
Yoshihisa Hagihara ◽  
Yutaka Kuroda
Keyword(s):  
Thermal Stress ◽  
Phosphate Buffer ◽  
Protein Concentration ◽  
Biochemical Analysis ◽  
Stress Condition ◽  
Binding Activity ◽  
Denatured State ◽  
E Coli ◽  
Structural And Functional Properties ◽  
Model Protein

In this study, we assessed the potential of arginine and lysine solubility-enhancing peptide (SEP) tags to control the solubility of a model protein, anti-EGFR VHH-7D12, in a thermally denatured state at a high temperature. We produced VHH-7D12 antibodies attached with a C-terminal SEP tag made of either five or nine arginines or lysines (7D12-C5R, 7D12-C9R, 7D12-C5K and 7D12-C9K, respectively). The 5-arginine and 5-lysine SEP tags increased the E. coli expression of VHH-7D12 by over 80%. Biophysical and biochemical analysis confirmed the native-like secondary and tertiary structural properties and the monomeric nature of all VHH-7D12 variants. Moreover, all VHH-7D12 variants retained a full binding activity to the EGFR extracellular domain. Finally, thermal stress with 45-minute incubation at 60 and 75 °C, where VHH-7D12 variants are unfolded, showed that the untagged VHH-7D12 formed aggregates in all of the four buffers, and the supernatant protein concentration was reduced by up to 35%. 7D12-C5R and 7D12-C9R did not aggregate in Na-acetate (pH 4.7) and Tris-HCl (pH 8.5) but formed aggregates in phosphate buffer (PB, pH 7.4) and phosphate buffer saline (PBS, pH 7.4). The lysine tags (either C5K or C9K) had the strongest solubilization effect, and both 7D12-C5K and 7D12-C9K remained in the supernatant. Altogether, our results indicate that, under a thermal stress condition, the lysine SEP tags solubilization effect is more potent than that of an arginine SEP tags, and the SEP tags did not affect the structural and functional properties of the protein.

Effect of weeds and nitrogen fertiliser on yield and grain protein concentration of wheat

1996 ◽  
Vol 36 (4) ◽  
pp. 443 ◽  
Author(s):  
MG Mason ◽  
RW Madin
Keyword(s):  
Grain Yield ◽  
Weed Control ◽  
Dry Matter ◽  
Protein Concentration ◽  
The Other ◽  
Grain Protein ◽  
Dry Matter Yield ◽  
Wheat Grain ◽  
Grain Protein Concentration ◽  
Nitrogen Fertiliser

Field trials at Beverley (19911, Salmon Gums (1991; 2 sites) and Merredin (1992; 2 sites), each with 5 rates of nitrogen (N) and 3 levels of weed control, were used to investigate the effect of weeds and N on wheat grain yield and protein concentration during 1991 and 1992. Weeds in the study were grasses (G) and broadleaf (BL). Weeds reduced both vegetative dry matter yield and grain yield of wheat at all sites except for dry matter at Merredin (BL). Nitrogen fertiliser increased wheat dry matter yield at all sites. Nitrogen increased wheat grain yield at Beverley and Merredin (BL), but decreased yield at both Salmon Gums sites in 1991. Nitrogen fertiliser increased grain protein concentration at all 5 sites-at all rates for 3 sites [Salmon Gums (G) and (BL) and Merredin (G)] and at rates of 69 kg N/ha or more at the other 2 sites [Beverley and Merredin (BL)]. However, the effect of weeds on grain protein varied across sites. At Merredin (G) protein concentration was higher where there was no weed control, possibly due to competition for soil moisture by the greater weed burden. At Salmon Gums (G), grain protein concentration was greater when weeds were controlled than in the presence of weeds, probably due to competition for N between crop and weeds. In the other 3 trials, there was no effect of weeds on grain protein. The effect of weeds on grain protein appears complex and depends on competition between crop and weeds for N and for water at the end of the season, and the interaction between the two.

scholarly journals Comparison Between Internal and External DSLs via RubyTL and Gra2MoL

2012 ◽  
pp. 109-131 ◽  
Author(s):  
Jesús Sánchez Cuadrado ◽  
Javier Luis Cánovas Izquierdo ◽  
Jesús García Molina
Keyword(s):  
Empirical Studies ◽  
Lessons Learned ◽  
The Other ◽  
Model Transformations ◽  
Domain Specific Languages ◽  
Model Driven ◽  
Domain Specific ◽  
Advantages And Disadvantages

Domain Specific Languages (DSL) are becoming increasingly more important with the emergence of Model-Driven paradigms. Most literature on DSLs is focused on describing particular languages, and there is still a lack of works that compare different approaches or carry out empirical studies regarding the construction or usage of DSLs. Several design choices must be made when building a DSL, but one important question is whether the DSL will be external or internal, since this affects the other aspects of the language. This chapter aims to provide developers confronting the internal-external dichotomy with guidance, through a comparison of the RubyTL and Gra2MoL model transformations languages, which have been built as an internal DSL and an external DSL, respectively. Both languages will first be introduced, and certain implementation issues will be discussed. The two languages will then be compared, and the advantages and disadvantages of each approach will be shown. Finally, some of the lessons learned will be presented.

scholarly journals Lessons Learned on the Other Side of Campus

2017 ◽  
Vol 81 (5) ◽  
pp. 81
Author(s):  
Lauren S. Schlesselman
Keyword(s):  
Lessons Learned ◽  
The Other

Moving In-Frame

2022 ◽  
pp. 217-234
Author(s):  
Andi Johnson ◽  
Richard Lessey ◽  
Rebeca Ramos O'Reilly ◽  
Jessica Shi
Keyword(s):  
Active Learning ◽  
Reflective Practice ◽  
Subject Area ◽  
Lessons Learned ◽  
The Other ◽  
Learning Engagement ◽  
Digital Education ◽  
The Subject ◽  
Further Development

The researchers explored the dual experience of individuals who are both taking dance and movement classes digitally at the same time as they are also teaching dance and movement classes digitally. By focusing on this duality through a series of interviews with practitioners, the researchers explore how the learner/educators do or do not adapt one set of skills into the other area of work and the lessons learned from this reflective practice. The results are analyzed and broken down into five sections: glitches of the practice, reflective practice, active learning, engagement, and reframing communication. Through further analysis, the researchers explore possibilities for shifting the mindset around digital education methods. The researchers then offer suggestions for further development in the field and where further research can expand on the subject area.

The Significant Impact of the COVID-19 Pandemic on Supply Chains

2022 ◽  
pp. 185-202
Author(s):  
Ana Paula Lopes
Keyword(s):  
Supply Chains ◽  
Consumer Demand ◽  
Lessons Learned ◽  
The Other ◽  
Operational Decisions ◽  
Demand And Supply ◽  
The World ◽  
Containment Measures ◽  
The Impact

As the COVID-19 pandemic has spread across the world, the existence of disruptions in demand and supply have become more severe, conducted by containment measures taken by countries and affecting different sectors around the world. Although businesses and workplaces are restarting activities in some countries, with containment measures gradually being lifted, overall consumer demand is expected to remain low, also determined by the loss of jobs and income. Therefore, the scale of the impact on supply chains exceeded anything most companies had anticipated. This study aims to understand how companies were affected and identify some lessons learned about their vulnerabilities and the possible ways to address them in the long term. On the other hand, it is intended to reveal some of the impacts of COVID-19 and make some practical suggestions that can help in political and operational decisions to strengthen and build additional resilience in supply chains in the future.

Darwin’s molecule

Hemoglobin ◽  
2018 ◽  
pp. 201-232
Author(s):  
Jay F. Storz
Keyword(s):  
Protein Function ◽  
Evolutionary Biology ◽  
Lessons Learned ◽  
Atomic Resolution ◽  
Molecular Adaptation ◽  
Genetic Adaptation ◽  
Biochemical Phenotype ◽  
Causal Connections ◽  
Model Molecule ◽  
Model Protein

Chapter 9 discusses conceptual issues in protein evolution and provides a synthesis of lessons learned from studies of hemoglobin function. Using hemoglobin as a model molecule, we can exploit an unparalleled base of knowledge about structure-function relationships and we can characterize biophysical mechanisms of molecular adaptation at atomic resolution. It is therefore possible to document causal connections between genotype and biochemical phenotype at an unsurpassed level of rigor and detail. Moreover, since the oxygenation properties of hemoglobin provide a direct link between ambient O2 availability and aerobic metabolism, genetically based changes in protein function can be related to ecologically relevant aspects of organismal physiology. We therefore have a solid theoretical framework for making predictions and for interpreting observed associations between biochemical phenotype and fitness-related measures of whole-animal physiological performance. The chapter explores case studies that illustrate how experimental research on functional properties of a well-chosen model protein can be used to address some of the most conceptually expansive questions in evolutionary biology: Is genetic adaptation predictable? Why does evolution follow some pathways rather than others?

Effect of Dimethyl Sulfoxide in Hydrophobic Modification of Cotton Filter Cloth by ARGET-ATRP Mechanism

2020 ◽  
Vol 993 ◽  
pp. 1407-1416 ◽  
Author(s):  
Zheng Li ◽  
Zi Jian He ◽  
Ying Cheng Zhou ◽  
Yi Tang ◽  
Yu Fang Chen ◽  
...  
Keyword(s):  
Dimethyl Sulfoxide ◽  
Crystal Form ◽  
Surface Roughening ◽  
Hydrophobic Modification ◽  
Filter Cloth ◽  
Water Contact ◽  
Grafting Reaction ◽  
Hydrophobically Modified ◽  
Arget Atrp ◽  
And Performance

In this paper, Dimethyl sulfoxide (DMSO) was used in the activating process of cotton filter cloth to improve its further hydrophobic modification reaction between cotton fabric and 1-octadecene via an electron transfer (ARGET) atom transfer radical polymerization (ATRP) mechanism. The major influences of DMSO on ARGET-ATRP process was discussed, and meanwhile, the microstructure changes, morphology feature and performance characteristics of cotton filter cloth during the reaction was explored by the SEM, AFM, EDS, XRD and TGA techniques.The result shows that DMSO can leads to cotton fibers adhesion and surface roughening under the ARGET-ATRP grafting reaction conditions, but has little changes on the crystal form, crystallinity and thermal properties of cellulose. At a DMSO dosage of 10%, the hydrophobically modified cotton filter cloth has a water contact angle (CA) of up to 141°. While naturally placed for 1 hour, the CA of hydrophobically modified cotton filter cloth can be stable at 116° with a decay rate of 17.5%, which proves that the hydrophobic stability of cotton filter cloth has been improved markedly. Furthermore, a better improvement for the hydrophobic stability of cotton filter cloth will significantly enhance the application of hydrophobic functional modified cellulosic materials.

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