Working towards the identification of virus-specific antibodies 

B cells, through their ability to produce specific antibodies, are essential to control invading viruses and play a primary role in the adaptive immune defense. Isolation of B cells from virus-infected individuals can serve as a first step towards the identification of virus-specific antibodies. 

Magnetic B cell separation with MACS® Technology 

For the identification of virus-specific antibodies, B cell isolation is required. To find optimal donors,  plasma samples of SARS-COV-2 infected patients can be analyzed for antiviral-response (e.g. binding to viral antigen in ELISA). In the next step, B cells can be isolated directly from the patients’ blood or PBMCs.  
B Cell Isolation Kits enable you to isolate highly pure, untouched B cells and B cell subsets from human PBMCs. 

Find the best kit for your project: 

Isolated cellsIsolation Kit
Activated B cells, malignant B cells and plasma cellsPan B Cell Isolation Kit, human
Resting, immature and naive B cells, depleted CD43+ cellsB Cell Isolation Kit II human
CD27+ memory B cellsMemory B Cell Isolation Kit, human
IgM expressing memory B cellsIgM+ Memory B Cell Isolation Kit, human
IgG expressing memory B cellsIgG+ Memory B Cell Isolation Kit, human
Switched Memory B cellsSwitched Memory B Cell Isolation Kit, human
Naive B cells  
(Please note that double negative memory B cells will not be depleted with this kit) 
Naive B Cell Isolation Kit II, human

B cell isolation directly from blood products 

To save valuable time, B cells can be directly isolated from blood products with minimal hands-on time. Automation options and no requirement for density gradient centrifugation or erythrocyte lysis. StraightFrom® MicroBeads allow magnetic cell separation directly from human whole blood, buffy coat, leukoreduction system chambers (LRSCs) and leukopaks. CD19+ and CD138+ B cells can be purified using the autoMACS® Pro Separator, the MultiMACS™ Cell24 Separator Plus, or manual separators.
 

Before isolation
  After isolation

Separation of CD138+ cells from a whole blood sample spiked with U266 cells, using the StraightFrom® Whole Blood and Bone Marrow CD138 MicroBeads. Cells were stained with CD45-FITC, CD138-PE, and CD19-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, dead cells, and CD45 cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Sorting of antigen specific B cells using the MACSQuant® Tyto® 

MACSQunat Tyto

Read the interview: How the MACSQuant® Tyto® Cell Sorter is helping a Danish research team to develop a new COVID-19 treatment.

B cell culture  

To be able to perform neutralizing, binding and functionality assays, B cells are cultured after they were separated. Ex vivo cultivation is an attractive option to increase B cells numbers, as their frequency is usually quite low. 

To make your primary B cell culture easier, we have developed the B Cell Expansion Kit, human. The formulation offers the perfect solution to expand and activate human B cells from a variety of starting materials. This enables expansion of B cells that are fully functional and ready for any downstream application. 

Expansion principle

Expansion rates

Expansion rate  B  Cell Expansion Kit, human
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• Up to 10-fold expansion of B cells in vitro within 14 days 
• Standardized and reproducible system 

Phenotype of expanded B cells

Expanded B cells display an activated phenotype with a distribution of memory and plasma cells similar to the original fraction. The amounts of naive B cells decreased under all culture conditions on day 14. This is in line with reports describing that naive B cells have a shorter survival rate. This way all important subsets are cultured and resent for your desired analysis.

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• Similar distribution of plasma cells, naive, and memory B cells (switched and non -switched)  
• At day 14, both showed a reduction of the naive B cell subset 
• The amount of memory cells was similar in the expanded cell population  
 

Multiparameter flow analysis of B cells  

As the final step in many workflows, flow analysis plays an important part in various B cell applications. Our flow reagents can ensure that the data you generate on your flow cytometer is reliable and consistent:

  • REAfinity™ Recombinant Antibodies – recombinantly generated antibodies that ensure lot-to-lot consistency and eliminate background signal 
  • MACSplex™ Cytokine Kits – analysis of multiple soluble cytokines in one sample using any standard flow cytometer 
  • Cytokine Secretion Assay – Detection Kits – flow analysis of cytokine secretion at a detection level of one in a million 
  • Build your own B cell flow panel with our Flow Panel Builder  

Analyze antigen specificity and find your lead candidate with automated flow cytometry

Flow cytometry can be used identify antigen-specific antibodies in tissue culture supernatants. The MACSQuant® X Flow Cytometer can perform these assays in a fast, standardized, and automated manner. This generates high-content information, that is both reproducible and reliable. The MACSQuant X is equipped with a syringe needle capable of volumetric pipetting, thereby enabling precise counting from sample volumes starting at 5 µL. The instrument is fully automated in its calibration and compensation procedures, as well as it’s pre-set templates, and instrument settings, significantly reducing experiment setup and optimization time. 

Fast and reliable high-throughput 96- and 384-well plate 
processing.
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Fast and reliable high-throughput 96- and 384-well plate processing.

 

Analyze your samples with high-speed: 
• 384-well plate in less than 60 minutes  
• 96-well plates in 15 minutes 
 

 Reliable high-throughput flow cytometry:

MACSQuant X Flow Cytometer

Check out our complete B cell portfolio