The SARS-CoV-2 B Cell Analysis Kit, human, has been developed for the fast and easy detection of SARS-CoV-2–specific B cells by binding of the SARS-CoV-2 specific proteins to the respective antigen-specific B cell receptor (BCR) on B cells circulating in the peripheral blood of individuals who developed a B cell response against SARS-CoV-2 proteins. The kit contains a SARS-CoV-2 recombinant protein, fluorochrome-conjugated antibodies for the identification and phenotyping of the B cells (memory B cells and isotypes), and 7-AAD for the exclusion of dead and apoptotic cells. The optimized flow panel

Data and images for SARS-CoV-2 B Cell Analysis Kits, human

Figures

Figure 1

Comparison of spike-specific B cell detection in COVID-19 convalescents versus healthy controls.
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Figure 1

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Figure 1

Figure 2

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Gating strategy for the identification and quantification of spike-specific B cells including isotyping of IgG, IgA, and IgM.
PBMCs of COVID-19 convalescent and healthy donors were obtained. B cells have been enriched using the REAlease® CD19 MicroBead Kit, human. Cells were used cryopreserved in StemMACS™ Cryo-Brew.
Samples of label-free B cells were stained and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. Cell debris, doublets, and dead cells were excluded from the analysis based on scatter signals and 7-AAD fluorescence.

Figure 2

Gating strategy for the identification and quantification of spike-specific B cells including isotyping of IgG, IgA, and IgM.
PBMCs of COVID-19 convalescent and healthy donors were obtained. B cells have been enriched using the REAlease® CD19 MicroBead Kit, human. Cells were used cryopreserved in StemMACS™ Cryo-Brew.
Samples of label-free B cells were stained and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. Cell debris, doublets, and dead cells were excluded from the analysis based on scatter signals and 7-AAD fluorescence.

Specifications for SARS-CoV-2 B Cell Analysis Kits, human

Overview

The SARS-CoV-2 B Cell Analysis Kit, human, has been developed for the fast and easy detection of SARS-CoV-2–specific B cells by binding of the SARS-CoV-2 specific proteins to the respective antigen-specific B cell receptor (BCR) on B cells circulating in the peripheral blood of individuals who developed a B cell response against SARS-CoV-2 proteins. The kit contains a SARS-CoV-2 recombinant protein, fluorochrome-conjugated antibodies for the identification and phenotyping of the B cells (memory B cells and isotypes), and 7-AAD for the exclusion of dead and apoptotic cells. The optimized flow panel and protocol ensures an easy and efficient analysis of SARS-CoV-2–specific B cells.

Detailed product information

Background information

B cells, also known as B lymphocytes express, dependent on their maturation stage, the antigen-specific B cell receptor (BCR) on their surface or secrete antigen-specific antibodies. They are part of the adaptive immune system and are crucial to mount a humoral, long lasting sterile immunity. B cells also play a role as professional antigen-presenting cells and secrete cytokines. They circulate through the body via the peripheral blood and account for 2–10% of all lymphocytes. Mature B cells in blood express the pan B cell marker CD19 and most mature B cells except plasma cells express CD20 and CD22. Additionally, different subsets of memory B cells and plasma cells can be identified based on their expression of Ig isotypes (IgM, IgD, IgG, IgA, IgE)1. Antigen-specific B cells usually occur at a frequency less than 0.05% for a specific antigen, but their number can vary depending on the phase and type of antigen or immunization
2-4
.
The quantitative and qualitative analysis of antigen-specific B cells specifically recognizing and reacting towards a defined antigen provide important information to understand their function in various immunological situations. The presence of these cells indicate that an individual is mounting an adaptive response to the specific, infective pathogen or to an immunization containing that specific antigen
5,6
. Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) induces an adaptive immune response leading to SARS-CoV-2–specific immunoglobulin and SARS-CoV-2–specific memory B and memory T cells with variable persistency and antiviral efficacy
5-7
.
Analysis and enrichment of antigen-specific B cells can contribute to the understanding of the role of cellular and humoral response and thus to the protection from SARS-CoV-2 infection after a previous infection or immunization.
9

Applications

Positive selection of SARS-CoV-2–specific B cells from peripheral blood mononuclear cells (PBMCs) of individuals exposed to SARS-CoV-2 after enrichment using the REAleaseR CD19 MicroBead Kit. The several-fold enriched antigen-specific B cells could be used for:
  • expansion in culture
  • molecular analysis, e.g. B cell receptor (BCR) cloning and sequencing
  • single clone culture

Resources for SARS-CoV-2 B Cell Analysis Kits, human

References for SARS-CoV-2 B Cell Analysis Kits, human

Publications

  1. Perez-Andres, M. et al. (2010) Human peripheral blood B-cell compartments: a crossroad in B-cell traffic Cytometry B Clin Cytom. 78(1): 47-60
  2. Smith, M. J. et al. (2017) Detection and Enrichment of Rare Antigen-specific B Cells for Analysis of Phenotype and Function J. Vis. Exp. 120: 55382
  3. Leyendeckers, H. et al. (1999) Correlation analysis between frequencies of circulating antigen-specific IgG-bearing memory B cells and serum titers of antigen-specific IgG Eur. J. Immunol. 29(4): 1406-1417
  4. Ward, S. M. et al. (2008) Direct ex vivo evaluation of long-lived protective antiviral memory B cell responses against hepatitis B virus J. Infect. Dis. 198(6): 813-817
  5. Hartley, G. E. et al. (2020) Rapid generation of durable B cell memory to SARS-CoV-2 spike and nucleocapsid proteins in COVID-19 and convalescence Sci Immunol 5(54): eabf8891
  6. Gaebler, C. et al. (2021) Evolution of antibody immunity to SARS-CoV-2 Nature 591: 639-644
  7. Dan, J. et al. (2021) Immunological memory to SARS-CoV-2 assessed for up to 8 months after infection. Science 371: eabf4063
  8. Sakharkar, M. et al. (2021) Prolonged evolution of the human B cell response to SARS-CoV-2 infection Sci Immunol 6(56): eabg6916
  9. Wang, Z. et al. (2021) mRNA vaccine-elicited antibodies to SARS-CoV-2 and circulating variants bioRxiv - the preprint server for Biology : 2021.01.15.426911
  10. Mazzoni, A. et al. (2021) First dose mRNA vaccination is sufficient to reactivate immunological memory to SARS-CoV-2 in ex COVID-19 subjects medRxiv - the preprint server for health sciences (doi.org/10.1101/2021.03.05.21252590)

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