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

从人外周血单核细胞中单细胞分选 HBsAg 结合记忆 B 细胞并进行抗体克隆

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作者:Yunjiao Zhou, Zhenmi Liu, Zijun Wang, Qianqian Zhang, Christian T Mayer, Till Schoofs, Michel C Nussenzweig, Ype P de Jong, Qiao Wang

Abstract

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).

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