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Iranian Journal of Basic Medical Sciences، جلد ۲۳، شماره ۸، صفحات ۱۰۵۳-۱۰۵۸
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عنوان انگلیسی Bioinformatics prediction and experimental validation of VH antibody fragment interacting with Neisseria meningitidis factor H binding protein
چکیده انگلیسی مقاله Objective(s): We previously conducted an in silico research on the interactions between the ribosome display-selected single chain variable fragment (scFv) and factor H binding protein (fHbp) of Neisseria meningitidis. We found that heavy chain variable (VH) fragment of this scFv had considerable affinity to fHbp. These results led us to evaluate the ability of this small antibody fragment in binding and detection of fHbp antigen.Materials and Methods: In this study, at first, the three-dimensional structure of VH fragment was simulated by Kotai Antibody Builder web server. By using ClusPro 2.0 web server, the 3D structure of the soluble form of fHbp (PDB: 2KC0) was docked to the modeled VH fragment to extract the structure of the complex’s binding. Molecular dynamics (MD) simulation was carried out using GROMACS 4.5.3 package for 65 ns. Secondly, coding sequence of VH fragment was cloned separately and expressed in Escherichia coli. After purification of the VH fragment, its binding activity to fHbp protein was analyzed by enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) method.Results: Important amino acids involved in antigen- antibody interaction were identified by analyzing the fHbp-VH complex. The ability of the VH antibody fragment to bind and detect fHbp antigen has been confirmed by the results of in silico analysis, ELISA and SPR methods.Conclusion: These results showed that this small fragment of antibody could be used for designing diagnostic kits.
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نویسندگان مقاله | Hediyeh Rafighdoust
Department of Biology, School of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran


| Shahrzad Ahangarzadeh
Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran


| Fatemeh Yarian
Cellular & Molecular Biology Research Center Shahid Beheshti University of Medical Sciences, Tehran, Iran|Department of Biotechnology, School of Advanced Technologies in Medicine Shahid Beheshti University of Medical Sciences, Tehran, Iran


| Ramezan Ali Taheri
Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences,Tehran, Iran


| Arezou Lari
Systems Biomedicine, Pasteur Institute of Iran, Tehran, Iran


| Mojgan Bandehpour
Cellular & Molecular Biology Research Center Shahid Beheshti University of Medical Sciences, Tehran, Iran


| Mona Salahshoor Dahr
Department of Biology, Faclty of Basic Science, Islamic Azad University Islamshahr Branch, Islamshahr, Iran
نشانی اینترنتی http://ijbms.mums.ac.ir/article_15834.html
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/87/article-87-2450582.pdf
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زبان مقاله منتشر شده en
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نوع مقاله منتشر شده Original Article
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