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

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 

作者机构：Nanfang HospitalSouthern Medical UniversityGuangzhouChina Institute of Genetic EngineeringSouthern Medical UniversityGuangzhouChina School of Traditional Chinese MedicineSouthern Medical UniversityGuangzhouChina School of Pharmaceutical ScienceSouthern Medical UniversityGuangzhouChina MOE/MOH Key Laboratory of Medical Molecular VirologyShanghai Medical CollegeFudan UniversityShanghaiChina Dgen Biotech Limited CompanyHong KongChina 

出 版 物：《Cellular & Molecular Immunology》 (中国免疫学杂志（英文版）)

年 卷 期：2012年第9卷第2期

页      面：184-190页

核心收录：

学科分类：0710[理学-生物学] 090602[农学-预防兽医学] 07[理学] 08[工学] 09[农学] 0906[农学-兽医学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

基　　金：supported by Dgen Biotech Ltd National Science and Technology Major Projects for ‘Major New Drugs Innovation and Development’ Guangdong Medical Research Foundation 

主　　题：antibody display antibody screening full-length antibody HBsAg-specific antibody mammalian display 

摘      要：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.