Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule

AbstractPseudomonas aeruginosa (PA) is a leading cause of nosocomial infections and death in cystic fibrosis patients. The study was conducted to evaluate the physicochemical structure, biological activity and serum stability of a recombinant anti-PcrV single chain variable antibody fragment genetically attached to the mCH3cc domain. The stereochemical properties of scFv-mCH3 (YFL001) and scFv (YFL002) proteins as well as molecular interactions towards Pseudomonas aeruginosa PcrV were evaluated computationally. The subcloned fragments encoding YFL001 and YFL002 in pET28a were expressed within the E. coli BL21-DE3 strain. After Ni–NTA affinity chromatography, the biological activity of the proteins in inhibition of PA induced hemolysis as well as cellular cytotoxicity was assessed. In silico analysis revealed the satisfactory stereochemical quality of the models as well as common residues in their interface with PcrV. The structural differences of proteins through circular dichroism spectroscopy were confirmed by NMR analysis. Both proteins indicated inhibition of ExoU positive PA strains in hemolysis of red blood cells compared to ExoU negative strains as well as cytotoxicity effect on lung epithelial cells. The ELISA test showed the longer serum stability of the YFL001 molecule than YFL002. The results were encouraging to further evaluation of these two scFv molecules in animal models.

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Improved biological activity of a single chain antibody fragment against human epidermal growth factor receptor 2 (HER2) expressed in the periplasm of Escherichia coli

Protein Expression and Purification ◽

10.1016/j.pep.2015.07.005 ◽

2015 ◽

Vol 116 ◽

pp. 66-74 ◽

Cited By ~ 4

Author(s):

Vajihe Akbari ◽

Hamid Mir Mohammad Sadeghi ◽

Abbas Jafarian-Dehkordi ◽

Daryoush Abedi ◽

C. Perry Chou

Keyword(s):

Escherichia Coli ◽

Epidermal Growth Factor Receptor ◽

Biological Activity ◽

Growth Factor ◽

Growth Factor Receptor ◽

Antibody Fragment ◽

Single Chain ◽

Single Chain Antibody ◽

Epidermal Growth ◽

Single Chain Antibody Fragment

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Bacterial expression of a single-chain antibody fragment (SCFV) that neutralizes the biological activity of human interferon

Molecular Immunology ◽

10.1016/0161-5890(93)90003-t ◽

1993 ◽

Vol 30 (9) ◽

pp. 805-812 ◽

Cited By ~ 10

Author(s):

Guy Froyen ◽

Isabelle Ronsse ◽

Alfons Billiau

Keyword(s):

Biological Activity ◽

Antibody Fragment ◽

Bacterial Expression ◽

Human Interferon ◽

Single Chain ◽

Single Chain Antibody ◽

Single Chain Antibody Fragment

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Generation and Evaluation of Chicken IgY-scFv for the Purposes of CPV Diagnosis and Therapy

10.21203/rs.3.rs-28773/v1 ◽

2020 ◽

Author(s):

Shikun Ge ◽

Long Xu ◽

Ben Li ◽

Fagang Zhong ◽

XiaoYing Zhang

Keyword(s):

Development Strategy ◽

Antibody Fragment ◽

Elisa Test ◽

Cross Reactivity ◽

Antibody Engineering ◽

Clinical Samples ◽

Similar Reaction ◽

Single Chain ◽

Diagnosis And Therapy ◽

T7 Phage Display

Abstract Background: Canine parvovirus (CPV) can cause acute and highly contagious enteritis in dog, antibodies have been used for diagnosis and therapy of CPV disease, generation of functional antibody fragments by using novel antibody engineering platforms is promising in veterinary practice. Results: The IgY single chain fragment variables (scFv) were generated by T7 phage display system and expressed in E. coli system after immunizing hens with virus-like particles (VLP) of CPV-VP2. The titer of the primary scFv library reached to 1.5×10 6 pfu/mL, and 95% of the phages contained the target fragments. The CPV-VLP and CPV-VP2 protein showed similar reaction values to the purified scFv in the ELISA test, and the results of ELISA analysis using IgY-scFv toward CPV clinical samples were consistent with commercial immunochromatographic assay (ICA) and PCR detection, the scFv did not show cross reactivity with canine distemper virus (CDV) and canine coronavirus (CCV). IgY-scFv was successfully expressed in CRFK cells, and in the virus suppression assay, 55% of CPV infections were eliminated within 24 hours. Docking results demonstrated that the number of amino acids of the binding sides between scFv and VP2 were AA37 and AA40, respectively. Conclusions: This study revealed the feasibility of a novel functional antibody fragment development strategy by generating diversified avian IgY-scFv libraries towards the pathogenic target of interest for both detection and therapeutic purposes in veterinary medicine.

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Construction of a single chain Fv (scFv195) antibody fragment against the human acetylcholine receptor. Contribution of light chain amino acid residues in receptor recognition

Immunology Letters ◽

10.1016/s0165-2478(97)88354-x ◽

1997 ◽

Vol 56 (1-3) ◽

pp. 375-376

Author(s):

P Tsantili

Keyword(s):

Amino Acid ◽

Acetylcholine Receptor ◽

Light Chain ◽

Antibody Fragment ◽

Amino Acid Residues ◽

Single Chain ◽

Single Chain Fv ◽

Receptor Recognition

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Faculty Opinions recommendation of Safety and pharmacokinetics of an anti-PcrV PEGylated monoclonal antibody fragment in mechanically ventilated patients colonized with Pseudomonas aeruginosa: a randomized,double-blind, placebo-controlled trial.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717847851.793453164 ◽

2012 ◽

Author(s):

Jonathon Truwit

Keyword(s):

Monoclonal Antibody ◽

Pseudomonas Aeruginosa ◽

Controlled Trial ◽

Antibody Fragment ◽

Double Blind ◽

Mechanically Ventilated ◽

Mechanically Ventilated Patients ◽

Double Blind Placebo ◽

Ventilated Patients

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Complexes of Cu (II) with a-Ketoglutaric Acid and 1- (o-tolyl) Biguanide Synthesis, characterization and biological Activity

Revista de Chimie ◽

10.37358/rc.19.10.7605 ◽

2019 ◽

Vol 70 (10) ◽

pp. 3603-3610

Author(s):

Madalina Mihalache ◽

Cornelia Guran ◽

Aurelia Meghea ◽

Vasile Bercu ◽

Ludmila Motelica ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Pseudomonas Aeruginosa ◽

Antimicrobial Activity ◽

Antibacterial Activity ◽

Biological Activity ◽

Candida Albicans ◽

Antifungal Activity ◽

Copper Complexes ◽

Substrate Adhesion ◽

Inert Substrate

The three copper complexes having a-ketoglutaric acid (H2A) and 1- (o-tolyl) biguanide (TB) ligands have been synthesized and characterized. The proposed formulas for these complexes are: [Cu(TB)(HA)]Cl (C1), [Cu(TB)(HA)CH3COO]�H2O (C2) and [Cu(TB)(HA)](NO3) (C3) where HA represents deprotonated H2A. The complexes obtained were tested for antibacterial activity against Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853, antifungal activity on Candida albicans ATCC 10231 and antitumor activity on HeLa tumor cells. Due to the antitumor, antifungal, antimicrobial activity and inhibition of inert substrate adhesion, complexes synthesized could be used for potential therapeutic applications.

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