Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule
| Autor: | Hoda Jahandar, Farzaneh Barkhordari, Fatemeh Davami, Yeganeh Talebkhan, Leila Nematollahi, Samira Komijani, Reza Moazzami, Fakhrisadat Hosseini, Elham Rismani, Elham Bayat, Soroush Sardari, Soma Hosseini |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Pore Forming Cytotoxic Proteins
0301 basic medicine Circular dichroism Science Bacterial Toxins 030106 microbiology medicine.disease_cause Article law.invention 03 medical and health sciences Affinity chromatography law Cell Line Tumor medicine Humans Computer Simulation Pseudomonas Infections Cloning Molecular Cytotoxicity Antigens Bacterial Cross Infection Multidisciplinary biology Chemistry Pseudomonas aeruginosa Biological techniques Biological activity medicine.disease Recombinant Proteins Hemolysis Anti-Bacterial Agents Molecular Docking Simulation 030104 developmental biology Biochemistry biology.protein Recombinant DNA Medicine Antibody Biotechnology Half-Life Single-Chain Antibodies |
| Zdroj: | Scientific Reports, Vol 11, Iss 1, Pp 1-14 (2021) Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | Pseudomonas 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|