scholarly journals Combinatorial microfluidic droplet engineering for biomimetic material synthesis

2016 ◽  
Vol 2 (10) ◽  
pp. e1600567 ◽  
Author(s):  
Lukmaan A. Bawazer ◽  
Ciara S. McNally ◽  
Christopher J. Empson ◽  
William J. Marchant ◽  
Tim P. Comyn ◽  
...  
Keyword(s):  
General Strategy ◽  
Emulsion Droplet ◽  
Material Synthesis ◽  
Widespread Application ◽  
Carrier Fluid ◽  
Wide Range ◽  
In Vitro Protein Expression ◽  
Selection Of ◽  
Vitro Protein

Although droplet-based systems are used in a wide range of technologies, opportunities for systematically customizing their interface chemistries remain relatively unexplored. This article describes a new microfluidic strategy for rapidly tailoring emulsion droplet compositions and properties. The approach uses a simple platform for screening arrays of droplet-based microfluidic devices and couples this with combinatorial selection of the droplet compositions. Through the application of genetic algorithms over multiple screening rounds, droplets with target properties can be rapidly generated. The potential of this method is demonstrated by creating droplets with enhanced stability, where this is achieved by selecting carrier fluid chemistries that promote titanium dioxide formation at the droplet interfaces. The interface is a mixture of amorphous and crystalline phases, and the resulting composite droplets are biocompatible, supporting in vitro protein expression in their interiors. This general strategy will find widespread application in advancing emulsion properties for use in chemistry, biology, materials, and medicine.

A PMMA microfluidic droplet platform for in vitro protein expression using crude E. coli S30 extract

Lab on a Chip ◽  
2009 ◽  
Vol 9 (23) ◽  
pp. 3391 ◽  
Author(s):  
N. Wu ◽  
Y. Zhu ◽  
S. Brown ◽  
J. Oakeshott ◽  
T. S. Peat ◽  
...  
Keyword(s):  
Protein Expression ◽  
E Coli ◽  
S30 Extract ◽  
In Vitro Protein Expression ◽  
Vitro Protein

scholarly journals cDNA display coupled with next-generation sequencing for rapid activity-based screening: Comprehensive analysis of transglutaminase substrate preference

2021 ◽  
Author(s):  
Jasmina Damnjanović ◽  
Nana Odake ◽  
Jicheng Fan ◽  
Beixi Jia ◽  
Takaaki Kojima ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Substrate Preference ◽  
Stable Complex ◽  
Next Generation ◽  
Library Size ◽  
Binary Model ◽  
Wide Range ◽  
Generation Sequencing ◽  
Selection Of

AbstractcDNA display is an in vitro display technology based on a covalent linkage between a protein and its corresponding mRNA/cDNA, where a stable complex is formed suitable for a wide range of selection conditions. A great advantage of cDNA display is the ability to handle enormous library size (1012) in a microtube scale, in a matter of days. To harness its benefits, we aimed at developing a platform which combines the advantages of cDNA display with high-throughput and accuracy of next-generation sequencing (NGS) for the selection of preferred substrate peptides of transglutaminase 2 (TG2), a protein cross-linking enzyme. After the optimization of the platform by the repeated screening of binary model libraries consisting of the substrate and non-substrate peptides at different ratios, screening and selection of combinatorial peptide library randomized at positions -1, +1, +2, and +3 from the glutamine residue was carried out. Enriched cDNA complexes were analyzed by NGS and bioinformatics, revealing the comprehensive amino acid preference of the TG2 at targeted positions of the peptide backbone. This is the first report on the cDNA display/NGS screening system to yield comprehensive data on TG substrate preference. Although some issues remain to be solved, this platform can be applied to the selection of other TGs and easily adjusted for the selection of other peptide substrates and even larger biomolecules.

scholarly journals Selection of Promising Lactobacilli Antagonistic to Campylobacter Jejuni

2019 ◽  
Vol 9 (1) ◽  
pp. 3983-3986
Keyword(s):  
Lactobacillus Plantarum ◽  
Campylobacter Jejuni ◽  
New Technologies ◽  
Lactobacillus Reuteri ◽  
Antagonistic Activity ◽  
Bacterial Strains ◽  
Opportunistic Bacteria ◽  
Wide Range ◽  
Selection Of

This article reflects the results of a study on the selection of promising lactobacilli antagonistic to Campilobacter jejuni, a strain that is the most common and more pathogenic for humans, carried out as part of a project to scientifically substantiate the use of new technologies in poultry feeding using special probiotic strains that increase productivity and obtaining poultry products of improved quality with the properties of functional food products. During the study, strains of lactic acid bacteria were obtained. The cultivation of strains of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus plantarum was carried out on liquid and agarized nutrient media MRS at 370С for 24 hours. In vitro antagonistic studies were performed using the two-way antagonistic method on a wide range of indicator crops. Since there is evidence of a specific mechanism for the manifestation of the antagonistic activity of lactobacilli to gram-negative and gram-positive bacteria, we used test strains of both groups of bacteria. The antagonistic activity of the studied cultures against pathogenic and opportunistic bacteria was determined by the zone of growth inhibition of indicator strains around the colonies of individual strains of lactobacilli and their consortium (in mm). Priority clinical isolates isolated from birds with intestinal infections were used as indicator cultures: Proteus vulgaris, Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Campylobacter jejuni and Campylobacter jejuni ATCC strains 33560. During the study, most bacterial strains of the genus Lactobacillus were highly antagonistic its activity against indicator strains. The most sensitive to the inhibitory effect of lactobacilli were E. coli, Campylobacter jejuni, S. typhimurium and P. vulgaris. The research results showed that the strain L.plantarum ATCC 8014 exhibits a more pronounced antagonistic activity than other strains of lactobacilli.

Application of in vitro protein solubility for selection of microalgae biomass as protein ingredient in animal and aquafeed

2020 ◽  
Vol 32 (6) ◽  
pp. 3955-3970
Author(s):  
G. Venkata Subhash ◽  
Neera Chugh ◽  
Supriya Iyer ◽  
Ashish Waghmare ◽  
Amar S. Musale ◽  
...  
Keyword(s):  
Protein Solubility ◽  
Protein Ingredient ◽  
Microalgae Biomass ◽  
Selection Of ◽  
Vitro Protein

scholarly journals Selection of Resistant Streptococcus pneumoniae during Penicillin Treatment In Vitro and in Three Animal Models

2003 ◽  
Vol 47 (8) ◽  
pp. 2499-2506 ◽  
Author(s):  
Jenny Dahl Knudsen ◽  
Inga Odenholt ◽  
Helga Erlendsdottir ◽  
Magnus Gottfredsson ◽  
Otto Cars ◽  
...  
Keyword(s):  
Animal Models ◽  
Mixed Culture ◽  
Observation Time ◽  
Treatment Regimens ◽  
Rabbit Tissue ◽  
Drug Concentrations ◽  
Wide Range ◽  
Different Strains ◽  
Selection Of

ABSTRACT Pharmacokinetic (PK) and pharmacodynamic (PD) properties for the selection of resistant pneumococci were studied by using three strains of the same serotype (6B) for mixed-culture infection in time-kill experiments in vitro and in three different animal models, the mouse peritonitis, the mouse thigh, and the rabbit tissue cage models. Treatment regimens with penicillin were designed to give a wide range of T>MICs, the amounts of time for which the drug concentrations in serum were above the MIC. The mixed culture of the three pneumococcal strains, 107 CFU of strain A (MIC of penicillin, 0.016 μg/ml; erythromycin resistant)/ml, 106 CFU of strain B (MIC of penicillin, 0.25 μg/ml)/ml, and 105 CFU of strain C (MIC of penicillin, 4 μg/ml)/ml, was used in the two mouse models, and a mixture of 105 CFU of strain A/ml, 104 CFU of strain B/ml, and 103 CFU of strain C/ml was used in the rabbit tissue cage model. During the different treatment regimens, the differences in numbers of CFU between treated and control animals were calculated to measure the efficacies of the regimens. Selective media with erythromycin or different penicillin concentrations were used to quantify the strains separately. The efficacies of penicillin in vitro were similar when individual strains or mixed cultures were studied. The eradication of the bacteria, independent of the susceptibility of the strain or strains or the presence of the strains in a mixture or on their own, followed the well-known PK and PD rules for treatment with β-lactams: a maximum efficacy was seen when the T>MIC was >40 to 50% of the observation time and the ratio of the maximum concentration of the drug in serum to the MIC was >10. It was possible in all three models to select for the less-susceptible strains by using insufficient treatments. In the rabbit tissue cage model, a regrowth of pneumococci was observed; in the mouse thigh model, the ratio between the different strains changed in favor of the less-susceptible strains; and in the mouse peritonitis model, the susceptible strain disappeared and was overgrown by the less-susceptible strains. These findings with the experimental infection models confirm the importance of eradicating all the bacteria taking part in the infectious process in order to avoid selection of resistant clones.

scholarly journals Phage Display Selection of an Anti-Idiotype-Antibody with Broad-Specificity to Deoxynivalenol Mycotoxins

Toxins ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 18
Author(s):  
Janne Leivo ◽  
Markus Vehniäinen ◽  
Urpo Lamminmäki
Keyword(s):  
In Vitro Selection ◽  
Specific Binding ◽  
Binding Properties ◽  
Phage Displays ◽  
Synthetic Antibody ◽  
Wide Range ◽  
Antibody Libraries ◽  
Rapid Generation ◽  
Selection Of

The use of synthetic antibody libraries and phage displays provides an efficient and robust method for the generation of antibodies against a wide range of targets with highly specific binding properties. As the in vitro selection conditions can be easily controlled, these methods enable the rapid generation of binders against difficult targets such as toxins and haptens. In this study, we used deoxynivalenol mycotoxin as a target to generate anti-idiotype-antibodies with unique binding properties from synthetic antibody libraries. The binding of the selected anti-idiotype antibodies can be efficiently inhibited with the addition of free isoforms of deoxynivalenol. The antibody was consecutively used to develop deoxynivalenol-specific ELISA and TRF-immunoassays, which can detect deoxynivalenol and two of the most common metabolic isoforms in the range of 78–115 ng/mL.

Proficient Target Selection in Structural Genomics by In Vitro Protein Expression on Gateway Recombination Plasmids

2002 ◽  
pp. 197-202 ◽  
Author(s):  
Vincent Monchois ◽  
Renaud Vincentelli ◽  
Céline Deregnaucourt ◽  
Chantal Abergel ◽  
Jean-Michel Claverie
Keyword(s):  
Protein Expression ◽  
Structural Genomics ◽  
Target Selection ◽  
In Vitro Protein Expression ◽  
Vitro Protein
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