ADCP Bioassay Effector Cell FcγRIIa (H Variant)/ NFAT Luciferase Reporter Jurkat Cell Line

Catalog #
71273
$6,875 *
Size: 2 vials
Qty
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Description

ADCP Bioassay Effector Cell FcγRIIa (H Variant)/NFAT Reporter Jurkat Cells are Jurkat T cells engineered to express firefly luciferase under the control of NFAT response elements, and human FcγRIIa, H variant. This cell line was functionally validated in a ADCP (antibody-dependent cell-mediated phagocytosis) assay.

Figure 1: Illustration of the mechanism of action of ADCP Bioassay Effector Cell FcγRIIa (H Variant)/NFAT Luciferase Reporter Jurkat Cell Line.
ADCP Bioassay Effector Cell FcγRIIa (H Variant)/NFAT Luciferase Reporter Jurkat cells are used as effector cells. The effector cells are co-cultured in the presence of target cells and an antibody of interest. The antibody binds to the target antigen on the target cell whereas its Fc portion binds to FcγRIIa on the cell surface of the effector cell, cross-linking the effector and target cells. Engagement of FcγRIIa leads to the activation of the NFAT pathway in the effector cells. Luciferase activity is proportional to the activation of the ADCP cascade.

Interested in screening and profiling inhibitors, blocking antibodies, or activators of FcγRIIa without the need to purchase and license the cell line? Check out our Cell Signaling Pathway Screening.

Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.

Synonyms
CD32a, FcgR2a
Product Info
Storage and Usage
Citations
Host Cell Line
Jurkat is a human leukemia cell line, Non-adherent T lymphocytes.
Supplied As
Each vial contains >1 x 106 cells in 1 ml of Cell Freezing Medium (BPS Bioscience #79796)
Materials Required But Not Supplied

Media Required for Cell Culture

Name Ordering Information
Thaw Medium 2 BPS Bioscience #60184
Growth Medium 2A BPS Bioscience #60190

 

Materials Required for Cellular Assays

Name Ordering Information
Anti-hHER2 human IgG1 Antibody R&D Systems #MAB9589-SP
Anti-CD20 Functional Antibody BPS Bioscience #71209
BT-474 Cells ATCC #HTB-20
Raji Cells ATCC #CCL-86
Human B Cells WIL2-S ATCC #CRL-8885
NFAT Reporter (Luc) – Jurkat Recombinant Cell Line BPS Bioscience #60621
96-well tissue culture-treated white clear-bottom assay plate  
One-Step™ Luciferase Assay System BPS Bioscience #60690
Luminometer  
UniProt #
P31995
Mycoplasma Testing

The cell line has been screened to confirm the absence of Mycoplasma species.

Background

Antibody-dependent cell-mediated phagocytosis (ADCP) is an important mechanism of action to consider during antibody drug development. FcγRIIa (also known as CD32a) is the predominant Fcγ receptor involved in the ADCP process. FcγRIIa is expressed in myeloid effector cells, including macrophages and neutrophils, where it plays a role in their activation. Engineered amino-acid substitutions in the Fc portion of monoclonal antibodies (mAb) can enhance the mAb-mediated phagocytosis of tumor cells by tumor-associated macrophages (TAMs). It is now clear that one of the major modes of action of therapeutic antibodies, that leads to positive outcomes, is their ability to trigger ADCP in TAMs. The binding of antibodies to Fcγ receptors leads to the phosphorylation of ITAMs (immunoreceptor tyrosine-based activation motifs) and activation of signaling pathways leading to phagocytosis via Rac-GEFs (guanine exchange factors). This mechanism of action has contributed to the success of rituximab (anti-CD20 antibody) in the treatment of chronic lymphocytic leukemia (CCL). Further understanding of this pathway, how to modulate and activate it, will lead to processes in cancer therapy.

References

Cao X., et al., 2022. Science Advances 8 (11): DOI: 10.1126/sciadv.abl9171