Flow cytometry analysis was performed as previously described.52 (link) PBMCs were isolated from blood of COVID-19 convalescent subjects using human lymphocyte separation medium. Cells were stained and mixed with fluorescent-labeled DNA-barcoded antigens and other fluorescent antibodies, and sorted by FACS with a MoFlo Astrios EQ Flow Cytometer (Beckman). The following antibodies were used to label the B cell subgroups: CD19-PB (BioLegend, Cat 302232), CD27-APC (BioLegend, Cat 302810), CD38-PE (BioLegend, Cat 303506). CD19+ CD27+ memory B cells and CD19+ CD27high CD38high plasma B cells were sorted for further scRNA-seq.
He B., Liu S., Wang Y., Xu M., Cai W., Liu J., Bai W., Ye S., Ma Y., Hu H., Meng H., Sun T., Li Y., Luo H., Shi M., Du X., Zhao W., Chen S., Yang J., Zhu H., Jie Y., Yang Y., Guo D., Wang Q., Liu Y., Yan H., Wang M, & Chen Y.Q. (2021). Rapid isolation and immune profiling of SARS-CoV-2 specific memory B cell in convalescent COVID-19 patients via LIBRA-seq. Signal Transduction and Targeted Therapy, 6, 195.
Corresponding Organization : Sun Yat-sen University
Other organizations :
Wuhan Institute of Virology, Chinese Academy of Sciences, Dongguan People’s Hospital, Sixth Affiliated Hospital of Sun Yat-sen University, Fifth Affiliated Hospital of Sun Yat-sen University, Third Affiliated Hospital of Sun Yat-sen University, Shanghai Medical College of Fudan University, Key Laboratory of Guangdong Province
Isolation of PBMCs from blood of COVID-19 convalescent subjects
dependent variables
Sorting of CD19+ CD27+ memory B cells and CD19+ CD27high CD38high plasma B cells by FACS
control variables
Use of human lymphocyte separation medium for PBMC isolation
Staining of cells with fluorescent-labeled DNA-barcoded antigens and other fluorescent antibodies
controls
No positive or negative controls were explicitly mentioned in the provided information.
Annotations
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