scholarly journals Improvement of High Affinity and Selectivity on Biosensors Using Genetically Engineered Phage by Binding Isotherm Screening

Viruses ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 248 ◽  
Author(s):  
Jong-Min Lee ◽  
Eun Choi ◽  
Juyun Park ◽  
Vasanthan Devaraj ◽  
ChunTae Kim ◽  
...  
Keyword(s):  
Screening Method ◽  
Target Material ◽  
Genetically Engineered ◽  
Titration Calorimetry ◽  
Binding Affinities ◽  
Binding Model ◽  
Screening Process ◽  
M13 Bacteriophage ◽  
Bacteriophage M13 ◽  
M13 Phage

The genetically engineered M13 bacteriophage (M13 phage), developed via directed evolutionary screening process, can improve the sensitivity of sensors because of its selective binding to a target material. Herein, we propose a screening method to develop a selective and sensitive bioreporter for toxic material based on genetically engineered M13 phage. The paraquat (PQ)-binding M13 phage, developed by directed evolution, was used. The binding affinities of the PQ-binding M13 phage to PQ and similar molecules were analyzed using isothermal titration calorimetry (ITC). Based on the isotherms measured by ITC, binding affinities were calculated using the one-site binding model. The binding affinity was 5.161 × 10−7 for PQ, and 3.043 × 10−7 for diquat (DQ). The isotherm and raw ITC data show that the PQ-binding M13 phage does not selectively bind to difenzoquat (DIF). The phage biofilter experiment confirmed the ability of PQ-binding M13 bacteriophage to bind PQ. The surface-enhanced Raman scattering (SERS) platform based on the bioreporter, PQ-binding M13 phage, exhibited 3.7 times the signal intensity as compared with the wild-type-M13-phage-coated platform.

M13 Bacteriophage Biolaminates for Nanomaterials with Improved Stiffness

2015 ◽  
Vol 1722 ◽  
Author(s):  
Christopher M. Warner ◽  
Amitabh Ghoshal ◽  
Michael F. Cuddy ◽  
Aimee R. Poda ◽  
Natalie D. Barker ◽  
...  
Keyword(s):  
Thin Films ◽  
High Performance ◽  
Liquid Crystalline ◽  
Genetically Engineered ◽  
Solution Chemistry ◽  
Thin Layers ◽  
M13 Bacteriophage ◽  
Material Development ◽  
Film Fabrication ◽  
M13 Phage

ABSTRACTIn nature, biomolecules guide the formation of hierarchically-ordered, lightweight, inorganic-organic composites such as corals, shells, teeth and bones. M13 bacteriophage has been used to mimic bio-inspired material development due to its rigid, nanoscale rod-like morphology. Liquid-crystalline monolayers of genetically engineered phage have been used to template crystallization of thin layers of inorganic and metallic materials. We have created thin films composed of engineered M13 phage capable of binding inorganic components. We employed both a dip-cast and a drop-cast film fabrication method on both smooth and rough gold, silica and glass casting surfaces to create thin films and 3D structures of various degrees of hierarchical order. We have found the engineered M13 phage and the inorganic mineral significantly affected both film morphology and the mechanical properties of the film. Similarly, film fabrication parameters such as solution chemistry, temperature, and pulling speed affected film properties. Using a calcium phosphate biomineralized 4E phage, film thickness increased linearly with the number of layers/dips in the phage solution. The stiffness of these composites (Young's modulus) were >80 GPa for mineralized, multilayer films. These materials are an order of magnitude stiffer than the biological equivalent collagen. Stiffness, however, does not appear to increase in a multilayer film beyond a saturation point. Ultimately, we have developed a platform for phage-based bio-composites for developing high performance materials.

scholarly journals One-step synthesis of M13 phage-based nanoparticles and their fluorescence properties

RSC Advances ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 1367-1375
Author(s):  
Jing Yi Lai ◽  
Naoya Inoue ◽  
Chuan Wei Oo ◽  
Hideya Kawasaki ◽  
Theam Soon Lim
Keyword(s):  
Nanoparticle Synthesis ◽  
Fluorescence Properties ◽  
Attractive Alternative ◽  
Alternative Source ◽  
M13 Bacteriophage ◽  
One Step ◽  
Fluorescent Nanoparticle ◽  
M13 Phage

M13 bacteriophage is an attractive alternative source for fluorescent nanoparticle synthesis.

Overview of EAF Screening Results on the 900 MWe NPP French Fleet

2016 ◽  
Author(s):  
Thomas Métais ◽  
Nicolas Robert ◽  
Pierre Genette ◽  
Nicolas Etchegaray
Keyword(s):  
Power Plants ◽  
Screening Method ◽  
Experimental Tests ◽  
Degradation Process ◽  
Primary System ◽  
Transfer Coefficients ◽  
Screening Process ◽  
The Usa ◽  
License Renewal ◽  
The Impact

In the wake of numerous experimental tests carried out in air and also in a PWR environment, both abroad and in France, an update of the current thermal fatigue codification is underway in France. Proposals are currently being integrated in the RCC-M code [1]. In parallel, it is necessary to evaluate the impact of codification evolution on the RCS components. In the USA, such evaluations have already been implemented for license renewal to operate power plants beyond their initial 40 years of operation. In order to reduce the scope of the calculations to perform, a preliminary screening was carried out on the various areas of the primary system components: this screening is detailed in an EPRI report [2]. The output of this screening process is a list of locations that are most prone to EAF degradation process and it is on these zones only that detailed EAF calculations are carried out. In France, a similar approach was defined in the perspective of the fourth ten-year visit of the 900 MWe plants (VD4 900 MWe) so as to map out all the locations that are most impacted by EAF and hence concentrate the calculation effort on these specific areas for the VD4 900 MWe. In that respect, a specific methodology to evaluate the factor to account for environmental effects or Fen [3] based on correlations [4] for hot and cold shocks was established. These correlations use data that is readily accessible in transient description documents and stress reports such as temperature change, heat transfer coefficients, ramp duration and geometry. The need for these correlations is specific to the French context due to a need for a preliminary and yet precise idea of the overall impact of the modifications brought to the RCC-M code in fatigue before the VD4 900 MWe. This paper presents the results of the screening method that was applied to the whole RCS of the 900 MWe NPP fleet.

scholarly journals Efficacy of Experimental Mouth Rinses on Caries-Related Biofilms in vitro

2021 ◽  
Vol 2 ◽  
Author(s):  
Josiana Steiger ◽  
Olivier Braissant ◽  
Tuomas Waltimo ◽  
Monika Astasov-Frauenhoffer
Keyword(s):  
Growth Rate ◽  
Antimicrobial Agents ◽  
Growth Parameters ◽  
Screening Method ◽  
Maximum Growth ◽  
Maximum Growth Rate ◽  
Screening Process ◽  
Mouth Rinses ◽  
Amine Fluoride

This study assessed the efficacy of tin and Polyethylenglycol (PEG-3) tallow aminopropylamine in different concentrations on Streptococcus mutans (S. mutans) biofilms to establish a new screening process for different antimicrobial agents and to gain more information on the antibacterial effects of these agents on cariogenic biofilms. Isothermal microcalorimetry (IMC) was used to determine differences in two growth parameters: lag time and growth rate; additionally, reduction in active biofilms was calculated. Experimental mouth rinses with 400 and 800 ppm tin derived from stannous fluoride (SnF2) revealed results (43.4 and 49.9% active biofilm reduction, respectively) similar to meridol mouth rinse (400 ppm tin combined with 1,567 ppm PEG-3 tallow aminopropylamine; 55.3% active biofilm reduction) (p > 0.05), while no growth of S. mutans biofilms was detected during 72 h for samples treated with an experimental rinse containing 1,600 ppm tin (100% active biofilm reduction). Only the highest concentration (12,536 ppm) of rinses containing PEG-3 tallow aminopropylamine derived from amine fluoride (AmF) revealed comparable results to meridol (57.5% reduction in active biofilm). Lower concentrations of PEG-3 tallow aminopropylamine showed reductions of 16.9% for 3,134 ppm and 33.5% for 6,268 ppm. Maximum growth rate was significantly lower for all the samples containing SnF2 than for the samples containing control biofilms (p < 0.05); no differences were found between the control and all the PEG-3 tallow aminopropylamine (p > 0.05). The growth parameters showed high reproducibility rates within the treated groups of biofilms and for the controls; thus, the screening method provided reliable results.

Experimental and numerical evaluation of a genetically engineered M13 bacteriophage with high sensitivity and selectivity for 2,4,6-trinitrotoluene

2019 ◽  
Vol 17 (23) ◽  
pp. 5666-5670 ◽  
Author(s):  
Won-Geun Kim ◽  
Chris Zueger ◽  
Chuntae Kim ◽  
Winnie Wong ◽  
Vasanthan Devaraj ◽  
...  
Keyword(s):  
Quantum Mechanics ◽  
Numerical Evaluation ◽  
High Sensitivity ◽  
Genetically Engineered ◽  
Experimental Results ◽  
Sensor System ◽  
M13 Bacteriophage ◽  
Sensitivity And Selectivity

This study includes the experimental results of a sensitive M13 bacteriophage-based sensor system that are well matched with the quantum mechanics calculation.

Optimization of the Existing Screening Method by Introducing Fen Correlations

2015 ◽  
Author(s):  
T. Métais ◽  
T. Gilman ◽  
P. Genette ◽  
A. Chinthapalli
Keyword(s):  
Power Plants ◽  
Screening Method ◽  
Experimental Tests ◽  
Degradation Process ◽  
Detailed Calculation ◽  
Screening Process ◽  
The Usa ◽  
License Renewal ◽  
The Impact ◽  
Geometrical Dimensions

In the wake of numerous experimental tests carried out in air and also in a PWR environment, both abroad and in France, an update of the current fatigue codification is underway. Proposals are currently being formulated in France [1] [2] and discussions are taking place in the frame of a French working group involving EDF, AREVA and CEA. In parallel with these worldwide modification efforts, it is necessary to evaluate their impact on the NSSS components. In the USA, many such evaluations have already been implemented for license renewal to operate power plants beyond their initial 40 years of operation. In order to reduce the scope of the calculations to perform, a preliminary screening was carried out on the various areas of the primary loop: this screening is detailed in an EPRI report [3]. The output of this screening process is a list of locations that are most prone to EAF degradation process and it is on these zones only that detailed EAF calculations are carried out. In France, with the approaching fourth decennial inspection of the 900 MWe (VD4 900 MWe) power plants, EDF needs also to map out the impact of these updates to the RCC-M code before initiating detailed calculation efforts. The EPRI report was not applicable as such to the French plants due to domestic specificities and more particularly, a need for a more detailed Fen estimation. A method was therefore developed by EDF, peer-reviewed by SI with the main innovation being the introduction of correlations enabling the calculation of Fen on the basis of the geometrical dimensions and the information available in the transient document. This paper presents how these correlations were built and proposes to benchmark them with some existing sample case problems.

PURIFICATION OF LYSOZYME BY INTRINSICALLY SHIELDED HYDROGEL BEADS

2013 ◽  
Vol 27 (19) ◽  
pp. 1341028
Author(s):  
CONG LI ◽  
R. ZHANG ◽  
L. WANG ◽  
A. BOWYER ◽  
R. EISENTHAL ◽  
...  
Keyword(s):  
Binary Mixture ◽  
High Purity ◽  
High Selectivity ◽  
Egg White ◽  
Titration Calorimetry ◽  
Binding Affinities ◽  
Affinity Separation ◽  
Hydrogel Beads ◽  
Relative Binding ◽  
Very High

Macro-sized intrinsically shielded hydrogel beads have been prepared from BSA and CM-dextran grafted with CB using a technique based on freeze-thawing gelation method. The size of the beads lies in around 500 μm. Isothemal titration calorimetry (ITC) showed that the relative binding affinities of the lysozyme for CB, compared with BSA, at pH 3.0 was stronger than that at pH 7.4. They were employed for the affinity separation of lysozyme using chromatography column. Their adsorption capacity for lysozyme at pH 3.0 is higher than that at pH 9. In a binary mixture of lysozyme and ovalbumin, the beads showed very high selectivity toward lysozyme. Lysozyme of very high purity (> 93%) was obtained from a mixture of lysozyme and ovalbumin, and 85% from egg white solution. The results indicate that the macro-sized bead can be used for the separation, purification, and recovery of lysozyme in a chromatograph column.

scholarly journals Secondary Structural Preferences of Some Antibacterial Cyclooctapeptides in the Presence of Calcium(II)

2012 ◽  
Vol 2012 ◽  
pp. 1-10
Author(s):  
Tarshona Stevens ◽  
Nykia McNeil ◽  
Xiuli Lin ◽  
Maria Ngu-Schwemlein
Keyword(s):  
Antibacterial Activity ◽  
Absorption Band ◽  
Titration Calorimetry ◽  
Conformational Structure ◽  
Binding Affinities ◽  
Beta Sheet ◽  
Antibacterial Compounds ◽  
Weak Binding ◽  
Beta Sheet Structure ◽  
Structural Preferences

The purpose of this study is to understand the interactions of some antibacterial cationic amphipathic cyclooctapeptides with calcium(II) and their secondary structural preferences. The thermodynamic parameters associated with calcium(II) interactions, between the antibacterial active cyclooctapeptides (COP 1–6) and those that did not exhibit significant activities (COP 7–9), were studied by isothermal titration calorimetry. Calcium(II) binding in the absence and presence of micellar dodecylphosphocholine (DPC), a membrane mimicking detergent, was conducted by circular dichroism (CD). Both groups of cyclopeptides showed weak binding affinities for calcium(II) (Kb ca. 10−3 M−1). However, CD data showed that the antimicrobial peptides COP 1–6 adopted a twisted beta-sheet structure (positive CD absorption band at ca. 203 nm) in the presence of calcium(II) in micellar DPC. In contrast, COP 7–9, which lacked antibacterial activity, adopted a different conformational structure (negative CD absorption band at ca. 203 nm). These results indicate that these cyclopeptides could adopt secondary structural preferences in the presence of calcium(II) amidst a hydrophobic environment to elicit their antibacterial activity. These findings could be useful in facilitating the design of cyclopeptide derivatives that can adopt this beta-sheet-like secondary structure and, thereby, provide a useful molecular template for crafting antibacterial compounds.

Recommendations on measurement and analysis of results obtained on biological substances using isothermal titration calorimetry (IUPAC Technical Report)

2008 ◽  
Vol 80 (9) ◽  
pp. 2025-2040 ◽  
Author(s):  
Frederick P. Schwarz ◽  
Timm Reinisch ◽  
Hans-Jürgen Hinz ◽  
Avadhesha Surolia
Keyword(s):  
Isothermal Titration Calorimetry ◽  
Small Molecule ◽  
Binding Constants ◽  
Titration Calorimetry ◽  
Heat Of Mixing ◽  
Buffer Solution ◽  
Binding Model ◽  
Binding Isotherm ◽  
Analyte Solution ◽  
Sample Vessel

Isothermal titration calorimetry (ITC) is widely used to determine the thermodynamics of biological interactions including protein-protein, small molecule-protein, protein-DNA, small molecule-DNA, and antigen-antibody interactions. An ITC measurement consists of monitoring the transfer of heat between an analyte solution in a sample vessel and a reference solution in a reference vessel upon injection of a small aliquot of titrant solution into the sample vessel at a fixed ITC operating temperature. A binding isotherm is generated from the heat-transferred-per-injection data and values for the binding constants, the apparent binding enthalpies, and the apparent ratio of the amount of titrant to analyte for the binding reaction are then determined from fits of a binding model, whether it is a single site, identical multi-site, or an interacting multi-site binding model, to the binding isotherm. Prior to the fitting procedure, corrections should be made for contributions from extraneous heat of mixing determined separately from injections of the titrant into just the dialysate/buffer solution. Ultra-high binding constants, which cannot be directly determined from an ITC measurement, can be determined by a displacement ITC method where injections of the tight-binding titrant into a solution of a weaker-binding titrant-analyte complex displaces the weaker-binding titrant from the complex. The Michaelis and catalytic constants can be determined for an enzyme reaction from injections of a substrate or enzyme titrant into an enzyme or substrate analyte solution. Several binding reactions are suggested to check the operating performance of the ITC. The reporting of ITC results must be specific with regard to the composition of the titrant and the analyte solutions, the temperature, and the model used in the analysis.

Biomimetic self-templating optical structures fabricated by genetically engineered M13 bacteriophage

2016 ◽  
Vol 85 ◽  
pp. 853-859 ◽  
Author(s):  
Won-Geun Kim ◽  
Hyerin Song ◽  
Chuntae Kim ◽  
Jong-Sik Moon ◽  
Kyujung Kim ◽  
...  
Keyword(s):  
Genetically Engineered ◽  
M13 Bacteriophage
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