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Development and Characterization of a Novel Single-Chain Antibody Against B-Cell Activating Factor

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Abstract

As a member of the tumor necrosis factor (TNF) superfamily, the B-cell activating factor (BAFF) plays a crucial role in B-cell survival and differentiation. Overexpression of this protein has been closely linked to autoimmune disorders and some B-cell malignancies. Using monoclonal antibodies (mAbs) against the BAFF soluble domain appears to be a complementary treatment for some of these diseases. This study aimed to produce and develop a specific Nanobody (Nb), a variable camelid antibody domain, against the soluble domain of BAFF protein. After camel immunization with recombinant protein and preparing cDNA from total RNAs separated from camel lymphocytes, an Nb library was developed. Individual colonies capable of binding selectively to rBAFF were obtained by periplasmic-ELISA, sequenced, and expressed in a bacterial expression system. The specificity and affinity of selected Nb were determined and its target identification and functionality were evaluated using flow cytometry.

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Acknowledgements

This study was financially supported by Pasteur Institute of Iran (Grant Number 994) and the National Institute for Medical Research Development (NIMAD), grant no. 943314.

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Authors and Affiliations

  1. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

    Rasoul Mardani-Jouneghani & Shiva Irani

  2. National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran

    Mahdi Habibi-Anbouhi

  3. Biotechnology Research Centre, Venom and Biotherapeutics Molecules Laboratory, Pasteur Institute of Iran, Tehran, 1316543551, Iran

    Rasoul Mardani-Jouneghani & Mahdi Behdani

  4. Zoonoses Research Centre, Pasteur Institute of Iran, Amol, Iran

    Mahdi Behdani

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  1. Rasoul Mardani-Jouneghani
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  2. Shiva Irani
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  3. Mahdi Habibi-Anbouhi
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Correspondence to Mahdi Behdani.

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Mardani-Jouneghani, R., Irani, S., Habibi-Anbouhi, M. et al. Development and Characterization of a Novel Single-Chain Antibody Against B-Cell Activating Factor. Mol Biotechnol 65, 1968–1978 (2023). https://doi.org/10.1007/s12033-023-00700-7

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