EasySep™ Human Memory B Cell Isolation Kit

Immunomagnetic cell isolation kit using particle release technology
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.
Advantages
• Highly purified human memory B cells isolated in as little as 33 minutes
• No-wash removal of EasySep™ Releasable RapidSpheres™
• Optional isolation of untouched naïve B cells from the same sample
Components
  • EasySep™ Human Memory B Cell Isolation Kit (Catalog #17864)
    • EasySep™ Human Memory B Cell Pre-Enrichment Cocktail, 2 x 1 mL
    • EasySep™ Dextran RapidSpheres™, 2 x 1 mL
    • EasySep™ Human CD27 Positive Selection Cocktail, 1 mL
    • EasySep™ Releasable RapidSpheres™, 2 x 1 mL
    • EasySep™ Release Buffer, 2 x 1 mL
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• Easy 50 EasySep™ Magnet (Catalog #18002)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
Subtype
Cell Isolation Kits
Cell Type
B Cells
Species
Human
Sample Source
PBMC
Selection Method
Positive
Application
Cell Isolation
Brand
EasySep
Area of Interest
Immunology
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. Y. Cao et al.
Abstract
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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.
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