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Computational analysis, design and expression of hybrid antibody in Single Chain Fragment Variable (scFv) form for identifying surface antigen factor H binding protein (fHbp) of Neisseria meningitidis
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Marjan Khajavi1    , Fatemeh Yarian 2, Shahrzad Ahangarzadeh3 , Mojgan Bandehpour4 , Mohsen Mohammadpour5 , Akram Jalali6 |
1- Msc Student, Department of Biology, School of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran
2- Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Fasa University of Medical Sciences, Fasa, Iran , f.yarian@fums.ac.ir
3- Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran
4- Cellular and Molecular Biology Research Center Shahid Beheshti University of Medical Sciences, Tehran, Iran
5- Department of Electrical and Computer Engineering, Fasa Branch, Islamic Azad University, Fasa, Iran
6- Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran |
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Abstract: (855 Views) |
Background: Meningococcal disease is an acute disease caused by the bacterium Neisseria meningitidis. The factor H binding protein (fHbp) virulence factor plays a vital role in the survival of the pathogen in the host. Single Chain Fragment Variable (scFv) antibodies are used for therapeutic and diagnostic applications. Utilizing computational study, Design, expression and affinity analysis of hybrid scFv for detecting the N. meningitidis were the main goals of this study.
Materials and methods: In this study, by performing a series of computational analysis (Antibody and Antigen modeling and Antigen-Antibody Docking), a hybrid antibody designed. The designed antibody has a high affinity to the fHbp protein. The hybrid antibody’s nucleotide was synthesized in the expression vector pET28(+a). Afterward, the protein expression in BL21 bacteria was considered. Subsequently, the protein purification was completed using Ni-NTA resin. Finally, its affinity to fHbp protein was checked using ELISA.
Results: VH1-VL2 form of hybrid antibody was selected with computational analysis. The SDS-PAGE and western blotting techniques were approved the expressed hybrid antibody. After purification of scFv and fHbp protein, in ELISA studies, the affinity of this scFv 7.6×10-9M was calculated.
Conclusion: Based on the hybrid antibody’s proper affinity and utilizing antibody engineering and computational analysis, other forms of single-chain antibodies can be produced. These products can be used for therapeutic and diagnostic purposes, as the appropriate hybrid affinity of the scFv |
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| Keywords: Hybrid single chain fragment antibody, Neisseria meningitidis, Factor H binding protein (fHbp), Cloning, Expression. |
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Full-Text [PDF 466 kb]
(595 Downloads)
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Semi-pilot: Basic |
Subject:
biotechnology
Received: 2022/07/9 | Accepted: 2022/10/16 | Published: 2023/03/30
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Khajavi M, Yarian F, Ahangarzadeh S, Bandehpour M, Mohammadpour M, Jalali A. Computational analysis, design and expression of hybrid antibody in Single Chain Fragment Variable (scFv) form for identifying surface antigen factor H binding protein (fHbp) of Neisseria meningitidis. MEDICAL SCIENCES 2023; 33 (1) :50-58 URL: http://tmuj.iautmu.ac.ir/article-1-2015-en.html
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