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Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning.

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机构: [1]Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai MedicalCollege, Fudan University, Shanghai 200032, China [2]West China School of Public Health, West China Hospital, Sichuan University, Chengdu 610041, China [3]Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA [4]Experimental Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health,Bethesda, MD 20892, USA [5]Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065, USA [6]Division of Gastroenterology and Hepatology, Weill Cornell Medicine, New York, NY 10065, USA7Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
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The isolation of human antibodies with naturally paired heavy and light chains is crucial for understanding the human antibody immune response. Here, we present a protocol for antibody cloning from the sorted single human memory B cells recognizing hepatitis B virus (HBV) S antigen (HBsAg). A two-fluorescent-dye labeling strategy against HBsAg allows for an improved sorting specificity, while non-relevant protein staining allows for the exclusion of non-specific B cells. This protocol could also be widely adapted for other antigens. For complete details on the use and execution of this protocol, please refer to Wang et al. (2020). © 2020 The Author(s).

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第一作者机构: [1]Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai MedicalCollege, Fudan University, Shanghai 200032, China
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通讯机构: [3]Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA [5]Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065, USA [6]Division of Gastroenterology and Hepatology, Weill Cornell Medicine, New York, NY 10065, USA7Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA
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