Production and evolution of a ScFv antibody for immunoassay of residual phenothiazine drugs in meat based on computational simulation

2017 ◽  
Vol 9 (30) ◽  
pp. 4455-4463 ◽  
Author(s):  
Fang Shu Shi ◽  
Lei Zhang ◽  
Wan Qiu Xia ◽  
Jing Liu ◽  
Hui Cai Zhang ◽  
...  
Keyword(s):  
Computational Simulation ◽  
Scfv Antibody ◽  
Directional Evolution ◽  
Phenothiazine Drugs

Production and directional evolution of a ScFv antibody based on computational simulation for immunoassay of phenothiazines in meat.

Virtual mutation and directional evolution of anti-amoxicillin ScFv antibody for immunoassay of penicillins in milk

2017 ◽  
Vol 517 ◽  
pp. 9-17 ◽  
Author(s):  
Xin He ◽  
Chang Fei Duan ◽  
Yong Hua Qi ◽  
Jun Dong ◽  
Geng Nan Wang ◽  
...  
Keyword(s):  
Scfv Antibody ◽  
Directional Evolution

scholarly journals Construction and directional evolution of anti-enrofloxacin ScFv antibody for the immunoassay of fluoroquinolones

2021 ◽  
Author(s):  
Fangyu Wang ◽  
Ning Li ◽  
Yunshang Zhang ◽  
Xuxefeng Sun ◽  
Yali Zhao ◽  
...  
Keyword(s):  
Amino Acid ◽  
Scfv Antibody ◽  
Phage Library ◽  
Enzyme Linked Immunosorbent Assay ◽  
Amino Acid Residues ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Relative Standard ◽  
Directional Evolution ◽  
Scfv Library

Abstract A recombinant anti-enrofloxacin single-chain antibody (scFv) was produced for the detection of enrofloxacin. An immunized mouse phage display scFv library with a capacity of 2.35×109 CFU/mL was constructed and used for anti-enrofloxacin scFv screening. After four rounds of bio-panning, 10 positives were isolated and identified successfully. The highest positive scFv was expressed in E. coli BL21. Then, its recognition mechanisms were studied using the molecular docking method. The result showed the amino acid residues Leu121 were the key residues for the binding of ScFv to ENR. Based on the results of virtual mutation, the ScFv antibody was evolved by directional mutagenesis of contact amino acid residue Leu121 to Asn. After the expression and purification, an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) based on the parental and mutant ScFv were established for enrofloxacin respectively. The IC50 value of the assay established with the ScFv mutant was 1.63 ng/mL, while the parental ScFv was 21.08 ng/mL, this result showed highly increased affinity with up to 12.9-folds improved sensitivity. The mean recovery for ENR ranged from 71.80% to 117.35% with 10.46% relative standard deviation between the intra-assay and the inter-assay. The results indicate that we have obtained a highly sensitive anti-ENR scFv by the phage library construction and directional evolution, and the scFv-based IC-ELISA is suitable for the detection of ENR residue in animal derived edible tissues and milk.

Production and Directional Evolution of Antisarafloxacin ScFv Antibody for Immunoassay of Fluoroquinolones in Milk

2016 ◽  
Vol 64 (42) ◽  
pp. 7957-7965 ◽  
Author(s):  
Jian Ping Wang ◽  
Jun Dong ◽  
Chang Fei Duan ◽  
Hui Cai Zhang ◽  
Xin He ◽  
...  
Keyword(s):  
Scfv Antibody ◽  
Directional Evolution

scholarly journals Construction and directional evolution of anti-enrofloxacin ScFv antibody for the immunoassay of fluoroquinolones

2021 ◽  
Author(s):  
Guangxu Xing ◽  
Yunshang Zhang ◽  
Fangyu Wang ◽  
Liuding Wen ◽  
Gaiping Zhang
Keyword(s):  
Amino Acid ◽  
Scfv Antibody ◽  
Phage Library ◽  
Enzyme Linked Immunosorbent Assay ◽  
Amino Acid Residues ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Relative Standard ◽  
Directional Evolution ◽  
Scfv Library

Abstract A recombinant anti-enrofloxacin single-chain antibody (scFv) was produced for the detection of enrofloxacin. An immunized mouse phage display scFv library with a capacity of 2.35×109 CFU/mL was constructed and used for anti-enrofloxacin scFv screening. After four rounds of bio-panning, 10 positives were isolated and identified successfully. The highest positive scFv was expressed in E. coli BL21. Then, its recognition mechanisms were studied using the molecular docking method. The result showed the amino acid residues Leu121 were the key residues for the binding of ScFv to ENR. Based on the results of virtual mutation, the ScFv antibody was evolved by directional mutagenesis of contact amino acid residue Leu121 to Asn. After the expression and purification, an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) based on the parental and mutant ScFv were established for enrofloxacin respectively. The IC50 value of the assay established with the ScFv mutant was 1.63 ng/mL, while the parental ScFv was 21.08 ng/mL, this result showed highly increased affinity with up to 12.9-folds improved sensitivity. The mean recovery for ENR ranged from 71.80% to 117.35% with 10.46% relative standard deviation between the intra-assay and the inter-assay. The results indicate that we have obtained a highly sensitive anti-ENR scFv by the phage library construction and directional evolution, and the scFv-based IC-ELISA is suitable for the detection of ENR residue in animal derived edible tissues and milk.

The study for constant force characteristic of C-shaped superelastic SMA

2020 ◽  
Vol 64 (1-4) ◽  
pp. 1253-1259
Author(s):  
Minghui Wang ◽  
Hongliu Yu
Keyword(s):  
Shape Memory Alloys ◽  
Shape Memory ◽  
Medical Device ◽  
Soft Tissues ◽  
Computational Simulation ◽  
Force Characteristic ◽  
Constant Force ◽  
Curve Shape ◽  
Artificial Sphincter ◽  
Round Bar

Clamping devices with constant force or pressure are desired in medical device, such as hemostatic forceps and the artificial sphincter, to prevent soft tissues from injures due to overloading. It is easily obtained by stretching an SMA wire. However, studies with SMA bending round bar have seldom been reported before. This paper studied constant force characteristic of C-shaped round bar with shape memory alloys. Optimization designs of the components were carried out with computational simulation. Numerical results show that the phenomenon of constant force strongly depends on contour curve shape and geometric dimensions of the C-shaped round bar of SMA component.

Influence of trees on the energy consumption of a social housing in mid-western Brazil

2019 ◽  
pp. 53-65
Author(s):  
Renata Domingos ◽  
Emeli Guarda ◽  
Elaise Gabriel ◽  
João Sanches
Keyword(s):  
Energy Consumption ◽  
Solar Radiation ◽  
Thermal Comfort ◽  
Air Conditioning ◽  
Computational Simulation ◽  
Building Energy ◽  
General Idea ◽  
Heat Island ◽  
Study Region ◽  
Ample Evidence

In the last decades, many studies have shown ample evidence that the existence of trees and vegetation around buildings can contribute to reduce the demand for energy by cooling and heating. The use of green areas in the urban environment as an effective strategy in reducing the cooling load of buildings has attracted much attention, though there is a lack of quantitative actions to apply the general idea to a specific building or location. Due to the large-scale construction of high buildings, large amounts of solar radiation are reflected and stored in the canyons of the streets. This causes higher air temperature and surface temperature in city areas compared to the rural environment and, consequently, deteriorates the urban heat island effect. The constant high temperatures lead to more air conditioning demand time, which results in a significant increase in building energy consumption. In general, the shade of the trees reduces the building energy demand for air conditioning, reducing solar radiation on the walls and roofs. The increase of urban green spaces has been extensively accepted as effective in mitigating the effects of heat island and reducing energy use in buildings. However, by influencing temperatures, especially extreme, it is likely that trees also affect human health, an important economic variable of interest. Since human behavior has a major influence on maintaining environmental quality, today's urban problems such as air and water pollution, floods, excessive noise, cause serious damage to the physical and mental health of the population. By minimizing these problems, vegetation (especially trees) is generally known to provide a range of ecosystem services such as rainwater reduction, air pollution mitigation, noise reduction, etc. This study focuses on the functions of temperature regulation, improvement of external thermal comfort and cooling energy reduction, so it aims to evaluate the influence of trees on the energy consumption of a house in the mid-western Brazil, located at latitude 15 ° S, in the center of South America. The methodology adopted was computer simulation, analyzing two scenarios that deal with issues such as the influence of vegetation and tree shade on the energy consumption of a building. In this way, the methodological procedures were divided into three stages: climatic contextualization of the study region; definition of a basic dwelling, of the thermophysical properties; computational simulation for quantification of energy consumption for the four facade orientations. The results show that the façades orientated to north, east and south, without the insertion of arboreal shading, obtained higher values of annual energy consumption. With the adoption of shading, the facades obtained a consumption reduction of around 7,4%. It is concluded that shading vegetation can bring significant climatic contribution to the interior of built environments and, consequently, reduction in energy consumption, promoting improvements in the thermal comfort conditions of users.

Deinking of recycled paper by coupling of the enzyme endo-β-1,4-D-glucanasa and the cellulose, and by using a laboratory flotation column

MRS Advances ◽  
2020 ◽  
Vol 5 (52-53) ◽  
pp. 2669-2678
Author(s):  
Jeovani González P. ◽  
Ramiro Escudero G
Keyword(s):  
Amino Acids ◽  
Sodium Hydroxide ◽  
Paper Industry ◽  
Computational Simulation ◽  
Cellulose Fibers ◽  
Residual Water ◽  
Chemical Reagent ◽  
Recycled Paper ◽  
Flotation Column ◽  
Hydrolysis Of

AbstractDeinking of recycled office (MOW) paper was carried out by using a flotation column and adding separately sodium hydroxide, and the enzyme Cellulase Thricodema Sp., as defibrillators.The de-inked cellulose fibers were characterized according to the standards of the paper industry, to compare the efficiency of the deinking of each chemical reagent used to hydrolyze the fibers and defibrillate them.The computational simulation of the molecular coupling between the enzyme and cellulose was performed, to establish the enzyme-cellulose molecular complex and then to identify the principal amino-acids of endo-β-1,4-D-glucanase in this molecular link, which are responsible for the hydrolysis of the cellulose.Experimental results show the feasibility to replace sodium hydroxide with the enzyme Cellulase Thricodema Sp., by obtaining deinked cellulose with similar optical and physical properties.The use of the enzyme instead of sodium hydroxide avoids the contamination of the residual water; in addition to that, the column is operated more easily, taking into consideration that the pH of the system goes from alkaline to neutral.

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