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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: (609 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]
(291 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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| References |
1. Read RC. Neisseria meningitidis and meningococcal disease: recent discoveries and innovations. Curr Opin Infect Dis 2019;32:601-8.
2. Booy R, Gentile A, Nissen M, Whelan J, Abitbol V. Recent changes in the epidemiology of Neisseria meningitidis serogroup W across the world, current vaccination policy choices and possible future strategies. Hum Vaccin Immunother 2019;15:470-80.
https://doi.org/10.1080/21645515.2018.1532248
3. Gasparini R, Amicizia D, Lai PL, Panatto D. Neisseria meningitidis, pathogenetic mechanisms to overcome the human immune defences. J Prev Med Hyg 2012;53:50-5.
4. Gasparini R, Amicizia D, Lai PL, Panatto D. Meningococcal B vaccination strategies and their practical application in Italy. J Prev Med Hyg 2015;56:E133-9.
5. McNeil LK, Zagursky RJ, Lin SL, Murphy E, Zlotnick GW, Hoiseth SK, et al. Role of factor H binding protein in Neisseria meningitidis virulence and its potential as a vaccine candidate to broadly protect against meningococcal disease. Microbiol Mol Biol Rev 2013;77:234-52.
https://doi.org/10.1128/MMBR.00056-12
6. Goulet DR, Atkins WM. Considerations for the Design of Antibody-Based Therapeutics. J Pharm Sci 2020;109:74-103.
https://doi.org/10.1016/j.xphs.2019.05.031
7. Chiu ML, Gilliland GL. Engineering antibody therapeutics. Curr Opin Struct Biol 2016;38:163-73.
https://doi.org/10.1016/j.sbi.2016.07.012
8. Bemani P, Mohammadi M, Hakakian A. ScFv Improvement Approaches. Protein Pept Lett 2018;25:222-9.
https://doi.org/10.2174/0929866525666171129225436
9. Yarian F, Kazemi B, Bandehpour M. Identification and characterization of a novel single-chain variable fragment (scFv) antibody against Neisseria meningitidis factor H-binding protein (fHbp). J Med Microbiol 2018;67:820-7.
https://doi.org/10.1099/jmm.0.000744
10. Bandehpour M, Yarian F, Ahangarzadeh S. Bioinformatics evaluation of novel ribosome display-selected single chain variable fragment (scFv) structure with factor H binding protein through docking. Journal of Theoretical and Computational Chemistry 2019;16:1750021.
https://doi.org/10.1142/S0219633617500213
11. Rafighdoust H, Ahangarzadeh S, Yarian F, Taheri RA, Lari A, Bandehpour M, et al. Bioinformatics prediction and experimental validation of VH antibody fragment interacting with Neisseria meningitidis factor H binding protein. Iran J Basic Med Sci 2020;23:1053-8.
12. Bandehpour M, Yarian F, Lari A, Farnia P. In silico evaluation of the interactions among two selected single chain variable fragments (scFvs) and ESAT-6 antigen of Mycobacterium tuberculosis. Journal of Theoretical and Computational Chemistry.2017;16:1750069.
https://doi.org/10.1142/S0219633617500699
13. Dunbar J, Krawczyk K, Leem J, Marks C, Nowak J, Regep C, et al. SAbPred: a structure-based antibody prediction server. Nucleic Acids Res 2016;44:W474-78.
https://doi.org/10.1093/nar/gkw361
14. Vajda S, Yueh C, Beglov D, Bohnuud T, Mottarella SE, Xia B, et al. New additions to the ClusPro server motivated by CAPRI. Proteins 2017;85:435-44.
https://doi.org/10.1002/prot.25219
15. Desta IT, Porter KA, Xia B, Kozakov D, Vajda S. Performance and Its Limits in Rigid Body Protein-Protein Docking. Structure 2020;28:1071-81.e3.
https://doi.org/10.1016/j.str.2020.06.006
16. Laskowski RA, Jabłońska J, Pravda L, Vařeková RS, Thornton JM. PDBsum: Structural summaries of PDB entries. Protein Sci 2018;27:129-34.
https://doi.org/10.1002/pro.3289
17. Yarian F, Bandehpour M, Seyed N, Kazemi B. Cloning, expression and purification of the factor H binding protein and its interaction with factor H. Iran J Microbiol 2016;8:29-35.
18. Byrne B, Stack E, Gilmartin N, O'Kennedy R. Antibody-based sensors: principles, problems and potential for detection of pathogens and associated toxins. Sensors 2009;9:4407-45.
https://doi.org/10.3390/s90604407
19. Rouphael NG, Stephens DS. Neisseria meningitidis: biology, microbiology, and epidemiology. Methods Mol Biol 2012;799:1-20.
https://doi.org/10.1007/978-1-61779-346-2_1
20. Payandeh Z, Rajabibazl M, Mortazavi Y, Rahimpour A, Taromchi AH, Dastmalchi S. Affinity maturation and characterization of the ofatumumab monoclonal antibody. J Cell Biochem 2019;120:940-50.
https://doi.org/10.1002/jcb.27457 [ DOI:10.1097/QCO.0000000000000606] |
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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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