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

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Cellular & Molecular Immunology 2012年第2期9卷 184-190页

作者： Chang-Zheng Li Zhong-Kun Liang Zhen-Rui Chen Hai-Bo Lou Ye Zhou Zhe-Huan Zhang Fei Yu Shuwen Liu Yuanping Zhou Shuguang Wu Wenling Zheng Wanlong Tan Shibo Jiang Chen Zhou Nanfang Hospital Southern Medical UniversityGuangzhouChina Institute of Genetic Engineering Southern Medical UniversityGuangzhouChina School of Traditional Chinese Medicine Southern Medical UniversityGuangzhouChina School of Pharmaceutical Science Southern Medical UniversityGuangzhouChina MOE/MOH Key Laboratory of Medical Molecular Virology Shanghai Medical CollegeFudan UniversityShanghaiChina Dgen Biotech Limited Company Hong KongChina

Hepatitis B immunoglobulin （HBIG） is important in the management of hepatitis B virus （HBV） infection. Aiming to develop recombinant monoclona# 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 FIp recombinase-mediated integration （FIp-ln） 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.

关键词： antibody display antibody screening full-length antibody HBsAg-specific antibody mammalian display

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