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Identification of HBsAg-specific antibodies from a mammalian cell displayed full-length human antibody library of healthy immunized donor

Cellular & Molecular Immunology volume 9, pages 184–190 (2012)Cite this article

Abstract

Hepatitis B immunoglobulin (HBIG) is important in the management of hepatitis B virus (HBV) infection. Aiming to develop recombinant monoclonal antibodies as an alternative to HBIG, we report the successful identification of HBV surface antigen (HBsAg)-specific antibodies from a full-length human antibody library displayed on mammalian cell surface. Using total RNA of peripheral blood mononuclear cells of a natively immunized donor as template, the antibody repertoire was amplified. Combining four-way ligation and the Flp recombinase-mediated integration (Flp-In) system, we constructed a mammalian cell-based, fully human, full-length antibody display library in which each cell displayed only one kind of antibody molecule. By screening the cell library using fluorescence-activated cell sorting (FACS), eight cell clones that displayed HBsAg-specific antibodies on cell surfaces were identified. DNA sequence analysis of the antibody genes revealed three unique antibodies. FACS data indicated that fluorescent strength of expression (FSE), fluorescent strength of binding (FSB) and relative binding ability (RBA) were all different among them. These results demonstrated that by using our antibody mammalian display and screening platform, we can successfully identify antigen-specific antibodies from an immunized full-length antibody library. Therefore, this platform is very useful for the development of therapeutic antibodies.

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Acknowledgements

This work was supported by Dgen Biotech Ltd (DGB12012008), National Science and Technology Major Projects for ‘Major New Drugs Innovation and Development’ (2011ZX09506–004) and Guangdong Medical Research Foundation (A2011363). The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.

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Author notes

  1. Chang-Zheng Li and Zhong-Kun Liang: These authors contributed equally to this work.

  2. Dr C Zhou, School of Pharmaceutical Science, Southern Medical University, 1838 North Guangzhou Road, Guangzhou 510515, China. E-mail: zyujun@yahoo.com

  3. Dr S Jiang, MOE/MOH Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, 130 Dongan Road, Shanghai 200032, China. E-mail: shibojiang@fudan.edu.cn

Authors and Affiliations

  1. Nanfang Hospital,, Guangzhou, China

    Chang-Zheng Li, Zhong-Kun Liang, Zhen-Rui Chen, Hai-Bo Lou, Zhe-Huan Zhang, Yuanping Zhou & Wanlong Tan

  2. Institute of Genetic Engineering, Southern Medical University, Guangzhou, China

    Chang-Zheng Li, Ye Zhou & Wenling Zheng

  3. School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China

    Chang-Zheng Li

  4. School of Pharmaceutical Science, Southern Medical University, Guangzhou, China

    Fei Yu, Shuwen Liu, Shuguang Wu & Chen Zhou

  5. MOE/MOH Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, Shanghai, China

    Shibo Jiang

  6. Dgen Biotech Limited Company, Hong Kong, China

    Chen Zhou

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  1. Chang-Zheng Li
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Li, CZ., Liang, ZK., Chen, ZR. et al. Identification of HBsAg-specific antibodies from a mammalian cell displayed full-length human antibody library of healthy immunized donor. Cell Mol Immunol 9, 184–190 (2012). https://doi.org/10.1038/cmi.2011.55

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  • DOI: https://doi.org/10.1038/cmi.2011.55

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