Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning
Summary: 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)....
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-12-01
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| Series: | STAR Protocols |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166720301167 |
| _version_ | 1818605983397576704 |
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| author | Yunjiao Zhou Zhenmi Liu Zijun Wang Qianqian Zhang Christian T. Mayer Till Schoofs Michel C. Nussenzweig Ype P. de Jong Qiao Wang |
| author_facet | Yunjiao Zhou Zhenmi Liu Zijun Wang Qianqian Zhang Christian T. Mayer Till Schoofs Michel C. Nussenzweig Ype P. de Jong Qiao Wang |
| author_sort | Yunjiao Zhou |
| collection | DOAJ |
| description | Summary: 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). |
| first_indexed | 2024-12-16T14:03:38Z |
| format | Article |
| id | doaj.art-5d4ee08913df4a6786a8fbd2879d1303 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-16T14:03:38Z |
| publishDate | 2020-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-5d4ee08913df4a6786a8fbd2879d13032022-12-21T22:28:58ZengElsevierSTAR Protocols2666-16672020-12-0113100129Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody CloningYunjiao Zhou0Zhenmi Liu1Zijun Wang2Qianqian Zhang3Christian T. Mayer4Till Schoofs5Michel C. Nussenzweig6Ype P. de Jong7Qiao Wang8Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, ChinaWest China School of Public Health, West China Hospital, Sichuan University, Chengdu 610041, ChinaLaboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USAKey Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, ChinaExperimental Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USALaboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USALaboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA; Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065, USA; Corresponding authorDivision of Gastroenterology and Hepatology, Weill Cornell Medicine, New York, NY 10065, USA; Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, USA; Corresponding authorKey Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166720301167 |
| spellingShingle | Yunjiao Zhou Zhenmi Liu Zijun Wang Qianqian Zhang Christian T. Mayer Till Schoofs Michel C. Nussenzweig Ype P. de Jong Qiao Wang Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning STAR Protocols |
| title | Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning |
| title_full | Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning |
| title_fullStr | Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning |
| title_full_unstemmed | Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning |
| title_short | Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning |
| title_sort | single cell sorting of hbsag binding memory b cells from human peripheral blood mononuclear cells and antibody cloning |
| url | http://www.sciencedirect.com/science/article/pii/S2666166720301167 |
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