Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule

Abstract 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 geneti...

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Autores principales:	Samira Komijani, Elham Bayat, Elham Rismani, Soma Hosseini, Reza Moazzami, Leila Nematollahi, Soroush Sardari, Yeganeh Talebkhan, Fatemeh Davami, Farzaneh Barkhordari, Fakhrisadat Hosseini, Hoda Jahandar
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fb0ca13a89d443c8b9cacb07341e3a89
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spelling	oai:doaj.org-article:fb0ca13a89d443c8b9cacb07341e3a892021-12-02T14:23:05ZCharacterization of a novel mCH3 conjugated anti-PcrV scFv molecule10.1038/s41598-021-86491-w2045-2322https://doaj.org/article/fb0ca13a89d443c8b9cacb07341e3a892021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-86491-whttps://doaj.org/toc/2045-2322Abstract 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.Samira KomijaniElham BayatElham RismaniSoma HosseiniReza MoazzamiLeila NematollahiSoroush SardariYeganeh TalebkhanFatemeh DavamiFarzaneh BarkhordariFakhrisadat HosseiniHoda JahandarNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Samira Komijani Elham Bayat Elham Rismani Soma Hosseini Reza Moazzami Leila Nematollahi Soroush Sardari Yeganeh Talebkhan Fatemeh Davami Farzaneh Barkhordari Fakhrisadat Hosseini Hoda Jahandar Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule
description	Abstract 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.
format	article
author	Samira Komijani Elham Bayat Elham Rismani Soma Hosseini Reza Moazzami Leila Nematollahi Soroush Sardari Yeganeh Talebkhan Fatemeh Davami Farzaneh Barkhordari Fakhrisadat Hosseini Hoda Jahandar
author_facet	Samira Komijani Elham Bayat Elham Rismani Soma Hosseini Reza Moazzami Leila Nematollahi Soroush Sardari Yeganeh Talebkhan Fatemeh Davami Farzaneh Barkhordari Fakhrisadat Hosseini Hoda Jahandar
author_sort	Samira Komijani
title	Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule
title_short	Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule
title_full	Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule
title_fullStr	Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule
title_full_unstemmed	Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule
title_sort	characterization of a novel mch3 conjugated anti-pcrv scfv molecule
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/fb0ca13a89d443c8b9cacb07341e3a89
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Characterization of a novel mCH3 conjugated anti-PcrV scFv molecule

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