Single-Cell Sorting of HBsAg-Binding Memory B Cells from Human Peripheral Blood Mononuclear Cells and Antibody Cloning.
STAR Protoc
; 1(3): 100129, 2020 12 18.
Article
in En
| MEDLINE
| ID: mdl-33377023
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).
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Flow Cytometry
/
Memory B Cells
Limits:
Humans
Language:
En
Journal:
STAR Protoc
Year:
2020
Document type:
Article
Affiliation country:
China