Erratum to “Virtual mutation and directional evolution of anti-amoxicillin ScFv antibody for immunoassay of penicillins in milk” [Anal. Biochem. 517 (2017) 9–17]

2017 ◽  
Vol 523 ◽  
pp. 44-45 ◽  
Author(s):  
Xin He ◽  
Chang Fei Duan ◽  
Yong Hua Qi ◽  
Jun Dong ◽  
Geng Nan Wang ◽  
...  
Keyword(s):  
Scfv Antibody ◽  
Directional Evolution

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 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.

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.

A Neutralization scFv Antibody against IL-1β Isolated from a NIPA-based Bacterial Display Library

2013 ◽  
Vol 14 (6) ◽  
pp. 571-581 ◽  
Author(s):  
Tianhe Li ◽  
Liming Xu ◽  
Guiping Ren ◽  
Chengkai Yin ◽  
Bing Zhou ◽  
...  
Keyword(s):  
Scfv Antibody ◽  
Bacterial Display ◽  
Bacterial Display Library

scholarly journals Isolation and characterizations of a novel recombinant scFv antibody against exotoxin A of Pseudomonas aeruginosa

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Zahra Shadman ◽  
Safar Farajnia ◽  
Mohammad Pazhang ◽  
Mohammadreza Tohidkia ◽  
Leila Rahbarnia ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Virulence Factors ◽  
High Reactivity ◽  
Scfv Antibody ◽  
Phage Library ◽  
Human Antibody ◽  
Exotoxin A ◽  
Phage Elisa ◽  
Human Scfv ◽  
Domain I

Abstract Background Pseudomonas aeruginosa is the leading cause of nosocomial infections, especially in people with a compromised immune system. Targeting virulence factors by neutralizing antibodies is a novel paradigm for the treatment of antibiotic-resistant pseudomonas infections. In this respect, exotoxin A is one of the most potent virulence factors in P. aeruginosa. The present study was carried out to identify a novel human scFv antibody against the P. aeruginosa exotoxin A domain I (ExoA-DI) from a human scFv phage library. Methods The recombinant ExoA-DI of P. aeruginosa was expressed in E. coli, purified by Ni-NTA column, and used for screening of human antibody phage library. A novel screening procedure was conducted to prevent the elimination of rare specific clones. The phage clone with high reactivity was evaluated by ELISA and western blot. Results Based on the results of polyclonal phage ELISA, the fifth round of biopanning leads to the isolation of several ExoA-DI reactive clones. One positive clone with high affinity was selected by monoclonal phage ELISA and used for antibody expression. The purified scFv showed high reactivity with the recombinant domain I and full-length native exotoxin A. Conclusions The purified anti-exotoxin A scFv displayed high specificity against exotoxin A. The human scFv identified in this study could be the groundwork for developing a novel therapeutic agent to control P. aeruginosa infections.

scholarly journals Chaperone-Assisted Soluble Expression of a Humanized Anti-EGFR ScFv Antibody in E. Coli

2015 ◽  
Vol 5 (Suppl 1) ◽  
pp. 621-627 ◽  
Author(s):  
Kamal Veisi ◽  
Safar Farajnia ◽  
Nosratollah Zarghami ◽  
Hamid Reza Khoram Khorshid ◽  
Nasser Samadi ◽  
...  
Keyword(s):  
Soluble Expression ◽  
Scfv Antibody ◽  
E Coli

Co-expression of a scFv antibody fragment and a reporter protein using lentiviral shuttle plasmid containing a self-processing furin-2A sequence

2013 ◽  
Vol 397 (1-2) ◽  
pp. 61-65 ◽  
Author(s):  
Sarah L. Appleby ◽  
Yazad Irani ◽  
Lauren A. Mortimer ◽  
Helen M. Brereton ◽  
Sonja Klebe ◽  
...  
Keyword(s):  
Antibody Fragment ◽  
Scfv Antibody ◽  
Reporter Protein ◽  
Shuttle Plasmid ◽  
Scfv Antibody Fragment
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