ADCP Bioassay Effector Cell FcγRIIa (H Variant)/ NFAT Luciferase Reporter Jurkat Cell Line
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.
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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 |
The cell line has been screened to confirm the absence of Mycoplasma species.
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.
Cao X., et al., 2022. Science Advances 8 (11): DOI: 10.1126/sciadv.abl9171