B cell isolation, expansion and analysis

Application notes, protocols and a scientific poster for B cell isolation, expansion and analysis

• MACSxpress® Technology allows isolation of B cells with high purity for sensitive and concordant microarray-based gene expression profiling
• Expansion of isolated B cells with the B Cell Expansion Kit, human
• Multicolor flow cytometric analysis of B cell subsets from mouse spleen
• A standardized and reproducible method for expansion and activation of human primary B cells 
• Gentle sorting of PBMC populations
• Increased sensitivity of flow cytometry for detection of minimal residual disease and circulating tumor cells in B Non-Hodgkin Lymphoma
• Positive Selection of Antibody Expressing Hybridoma Cells 

B cell separation

We have a diverse range of solutions for B cell separation. These solutions include untouched isolation, positive isolation, isolation directly from different blood products and REAlease® Technology, to offer you a streamlined method for fast and efficient B cell isolation. Whatever your subset of interest or starting material, we have the right tool for you.

MACS Technology offers a solution to obtain a highly purified population separated by positive selection or untouched cells from whole blood, untouched B cells and B cell subsets from human PBMCs. 

REAlease Technology allows for magnetic cell isolation with subsequent removal of any beads and labels from your B cells of interest. Clever utilization of recombinant antibody fragments ensures cell surface labeling in just a few steps.  

The technology relies on recombinantly engineered antibody fragments for cell surface labeling, ensuring highly reproducible results. The REAlease Biotin Complex binds to the target cells. Labeling of the REAlease Biotin Complex with Anti-Biotin MicroBeads promotes magnetic isolation of these cells. Following cell separation, both MicroBeads and the REAlease Biotin Complex are gently removed, leaving the cells bead- and label-free. Enriched cells are then suitable for magnetic relabeling and any application where label-free cells are essential. 

Positive separation of B cells from blood products using StraightFrom® MicroBeads

StraightFrom Technology ensures isolated B cells directly from blood products whilst saving time. The StraightFrom® MicroBeads provide magnetic cell separation directly from human blood products, including, 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.
 

Untouched B cell isolation from blood products using MACSxpress® Technology

For smaller starting material volumes the MACSxpress Technology isolates viable and functional B cells quickly and conveniently from blood and blood products. This cell isolation technology is based on micro-sized MACSxpress Beads, which allow the removal of non-target cells by immunomagnetic depletion using the MACSxpress Separator. All without the requirement of density gradient centrifugation or red blood cell (RBC) lysis. 
 

For the isolation of rare B cell sub-populations the The MACSQuant® Tyto® Cell Sorter is revolutionizing cell sorting. Our patented microchip-based technology opens up new possibilities with high-speed multiparameter cell sorting in the safety of a fully enclosed and sterile cartridge system, the MACSQuant Tyto Cartridge. This enables gentle sorting of B cells, leading to high target cell purity and viability of theses sensitive cells.
 

B cell culture 

Due to the low frequency of B cells in peripheral blood, ex vivo cultivation is an ideal option to increase B cell numbers; either to study their biology, or transform them and analyze their antibody production as well as specificity. Often, this can prove difficult, and many of the challenges associated with this can be overcome using our  B Cell Expansion Kit, human. The formulation offers the perfect solution to expand and activate human B cells from various sources. This enables expansion of B cells that are fully functional and ready for any downstream application.

Reliable activation and expansion of B cells is a clear requirement for effective downstream experiments. The B Cell Expansion Kit, human is optimized for primary B cell expansion in vitro. The B Cell Expansion Kit consist of a recombinant CD40L multimer, a cross-linking antibody, and IL-4. Cross-linking of CD40 expressed on B cells leads to cell activation and proliferation. Combining CD40L together with IL-4, which is a known growth factor for activated B cells, has been shown to be crucial for effective activation and proliferation of B cells.

Expanded B cells display an activated phenotype with a distribution of memory and plasma cells similar to the original fraction. The amount 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. 
 

Multiparameter flow analysis of B cells

As the final step in many workflows, flow analysis plays an important part in various B cell applications. The following flow reagents can help you make sure 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