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概要
The EasySep™ Human Memory B Cell Isolation Kit is a two-step method designed to isolate CD27+ B cells from fresh or previously frozen peripheral blood mononuclear cells. First, CD27+ cells are isolated by column-free immunomagnetic positive selection using antibody complexes and EasySep™ Releasable RapidSpheres™. Then, bound magnetic particles are removed from the EasySep™-isolated CD27+ cells, and unwanted non-B cells are targeted for depletion using antibody complexes and EasySep™ Dextran RapidSpheres™. The final isolated fraction contains highly purified CD27+ memory B cells that are immediately ready for downstream applications. An optional protocol allows for the isolation of naïve B cells in parallel for use in functional studies.
技术资料
| Document Type | 产品名称 | Catalog # | Lot # | 语言 |
|---|---|---|---|---|
| Product Information Sheet | EasySep™ Human Memory B Cell Isolation Kit | 17864 | All | English |
| Safety Data Sheet 1 | EasySep™ Human Memory B Cell Isolation Kit | 17864 | All | English |
| Safety Data Sheet 2 | EasySep™ Human Memory B Cell Isolation Kit | 17864 | All | English |
| Safety Data Sheet 3 | EasySep™ Human Memory B Cell Isolation Kit | 17864 | All | English |
| Safety Data Sheet 4 | EasySep™ Human Memory B Cell Isolation Kit | 17864 | All | English |
| Safety Data Sheet 5 | EasySep™ Human Memory B Cell Isolation Kit | 17864 | All | English |
数据及文献
Data
Starting with PBMCs, the memory B cell content (CD19+CD27+) of the isolated fraction is typically 97 ± 2% (mean ± SD using the purple EasySep™ Magnet). Using the optional protocol, the naïve B cell content (CD19+CD27-) of the isolated fraction is typically 93 ± 5% (mean ± SD).
Publications (1)
Cell 2020
Potent Neutralizing Antibodies against SARS-CoV-2 Identified by High-Throughput Single-Cell Sequencing of Convalescent Patients' B Cells.
Abstract
Abstract
The COVID-19 pandemic urgently needs therapeutic and prophylactic interventions. Here, we report the rapid identification of SARS-CoV-2-neutralizing antibodies by high-throughput single-cell RNA and VDJ sequencing of antigen-enriched B cells from 60 convalescent patients. From 8,558 antigen-binding IgG1+ clonotypes, 14 potent neutralizing antibodies were identified, with the most potent one, BD-368-2, exhibiting an IC50 of 1.2 and 15 ng/mL against pseudotyped and authentic SARS-CoV-2, respectively. BD-368-2 also displayed strong therapeutic and prophylactic efficacy in SARS-CoV-2-infected hACE2-transgenic mice. Additionally, the 3.8 {\AA} cryo-EM structure of a neutralizing antibody in complex with the spike-ectodomain trimer revealed the antibody's epitope overlaps with the ACE2 binding site. Moreover, we demonstrated that SARS-CoV-2-neutralizing antibodies could be directly selected based on similarities of their predicted CDR3H structures to those of SARS-CoV-neutralizing antibodies. Altogether, we showed that human neutralizing antibodies could be efficiently discovered by high-throughput single B cell sequencing in response to pandemic infectious diseases.
