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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.
We offer various antigens for the major immunogenic proteins of SARS-CoV-2 as well as the entry receptor ACE2. The antigens are available in different sizes with or without biotinylation. Find a comprehensive product overview and see how they were applied to detect SARS-CoV-2–specific antibodies in serum samples here!
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.
|Isolated cells||Isolation Kit|
|Activated B cells, malignant B cells and plasma cells||Pan B Cell Isolation Kit, human|
|Resting, immature and naive B cells, depleted CD43+ cells||B Cell Isolation Kit II human|
|CD27+ memory B cells||Memory B Cell Isolation Kit, human|
|IgM expressing memory B cells||IgM+ Memory B Cell Isolation Kit, human|
|IgG expressing memory B cells||IgG+ Memory B Cell Isolation Kit, human|
|Switched Memory B cells||Switched 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|
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.
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.
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.
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.
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:
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.
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