Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning
oleh: Yunjiao Zhou, Zhenmi Liu, Zijun Wang, Qianqian Zhang, Christian T. Mayer, Till Schoofs, Michel C. Nussenzweig, Ype P. de Jong, Qiao Wang
| Format: | Article |
|---|---|
| Diterbitkan: | Elsevier 2020-12-01 |
Deskripsi
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).