Development of specific nanobodies (VHH) for CD۱۹ immuno-targeting of human B-lymphocytes
Publish place: Iranian Journal of Basic Medical Sciences، Vol: 21، Issue: 5
Publish Year: 1397
نوع سند: مقاله ژورنالی
زبان: English
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شناسه ملی سند علمی:
JR_IJBMS-21-5_003
تاریخ نمایه سازی: 27 مهر 1400
Abstract:
Objective(s): CD۱۹ is a transmembrane glycoprotein of immunoglobulin superfamily. In order to treat lymphoma, monoclonal antibodies (mAb) can target different antigens, including CD۱۹, CD۲۰ and CD۲۲ on the surface of B-cells. Along with biotechnology progress, a new generation of antibodies is introduced, with the purpose of eliminating the defects of the previous generation. Among the most developed one are nanobodies (Nb). Nbs are a unique kind of camelid single domain antibody fragments with a broad range of medical applications. Unique physicochemical properties of Nbs have made them ideal candidates for therapeutic and diagnostic applications. Materials and Methods: An immune gene library was created, and several CD۱۹ specific Nbs were selected through antigen panning process, and their molecular properties as well as specificity, sensitivity, affinity and immunoreactivity against CD۱۹ positive and negative cells were evaluated. Results: The Nb library was prepared with ۷.۲ x۱۰۷ members. We managed to isolate a panel of CD۱۹-specific Nbs after the last round of selection with the affinity of isolated Nbs being estimated at the standard range of ۱۵-۳۵ nM. Sequence analysis of positive clones was indicative of the fact that ۱۲ variable sequences were confirmed. Of all these ۱۲ clones, ۲ clones with the greatest level signal in ELISA underwent subsequent analysis. Our sequencing results indicated high sequence homology (approximately ۹۰%) between the Nb and Homa variable immunoglobulin domains.Conclusion: Specific Nbs possess the potential to be used as novel therapeutic approaches in order to treat autoimmune diseases and B-cell lymphoma.
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Authors
Seyed Reza Banihahsemi
Department of Medical Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Ahmad Zavaran Hosseini
Department of Medical Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Fatemeh Rahbarizadeh
Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Davoud Ahmadvand
Department of Medical Biochemistry, Faculty of Applied Medical Sciences, Iran University of Medical Sciences,Tehran, Iran
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