Human EGFR-expressing Raji Cells
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Human lymphoblast cells - ADCC EGFR Target Cells
3-7 x 10^6 cells
Human EGFR-expressing B cells
Raji-hEGFR cells were developed from the human Raji cell line, a human B lymphocyte-derived cell line, and engineered to stably overexpress the human EGFR gene. Raji cells have been successfully used as target cells in human effector studies such as antibody-dependent cellular cytotoxicity (ADCC), either with peripheral blood mononuclear cells, natural killer (NK) cells, or T cell-derived Jurkat reporter cells.
Epidermal growth factor receptor (EGFR), also known as HER1 or ERBB1, is a member of the EGFR/ERBB family of receptor tyrosine kinases [1,2]. Under normal physiological conditions, activation of EGFR by various ligands, including epidermal growth factor (EGF) and transforming growth factor (TGF)–α, leads to the formation of an active EGFR-homodimer or heterodimer with another member of the ERBB family such as HER2. EGFR-dependent signaling is crucial in the regulation of tissue development and homeostasis . However, uncontrolled surface overexpression and/or enhanced signaling activity of EGFR is a major driver of tumor progression (e.g. lung, breast, and glioblastoma) [1,2]. Thus, it is considered an important tumor-associated antigen and oncogene. EGFR-targeting monoclonal antibody (mAb) therapies (e.g. Cetuximab) have been approved for the treatment of a variety of cancers [1,2]. EGFR remains an important therapeutic target for the development of more potent mAbs (e.g. increased effector function).
Features of Raji-hEGFR cells:
- Stable overexpression of the human EGFR gene
- Characterized by a number of cell-surface expressed markers including the B cell receptor (BCR), CD19, and CD20
- Constitutive expression of various immune checkpoints (ICs) such as CD27, CD70, CD80, PD-L1, and 4-1BBL
Applications for Raji-hEGFR cells:
- Target cell line for ADCC assays using InvivoGen's Jurkat-Lucia™ NFAT-CD16 cells
- Target cell line for cell toxicity assays using NK or CAR-T cells
- For use in the development of other functional assays
Validation of Raji-hEGFR cells:
- Overexpression of EGFR verified by flow cytometry
- Functionally tested as target cells in ADCC assays using anti-human EGFR mAbs and Jurkat-Lucia™ NFAT-CD16 cells
- Guaranteed mycoplasma-free
1. Sigismund, S. et al. 2018. Emerging functions of the EGFR in cancer. Mol Oncol 12, 3-20.
2. Sabbah, D.A. et al. 2020. Epidermal Growth Factor Receptor (EGFR) Structure, Signaling Pathways, Interactions, and Recent Updates of EGFR Inhibitors. Curr Top Med Chem 20, 815-834.
Validation of the expression of human EGFR by Raji-hEGFR cells. Raji-Null (A) and Raji‑hEGFR (B) cells were incubated with a PE-conjugated Anti-hEGFR mAb for 30 minutes. The binding affinity was then measured using flow cytometry.
Comparison of ADCC potency for native and engineered anti-human EGFR antibody isotypes. Raji-hEGFR cells were incubated with gradient concentrations of Anti-hEGFR or Anti-β-galactosidase (β-Gal) mAbs for 1 hour. Jurkat-Lucia™ NFAT-CD16 effector cells were then co-incubated with target cells for 6 hours. NFAT activation, reflecting the induced ADCC response, was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™. Percentages of the maximal response normalized to the IgG1 isotype are shown.
Antibiotic resistance: Blasticidin
Test medium: IMDM, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated FBS, Pen-Strep (100 U/ml-100 µg/ml)
- Human EGFR expression has been verified by flow-cytometry.
- Induction of antibody-dependent cellular cytotoxicity (ADCC) has been validated using anti-hEGFR antibodies and InvivoGen's Jurkat-NFAT Lucia™ CD16 reporter cell line.
- The stability for 20 passages following thawing has been verified.
- Raji-hEGFR cells are guaranteed mycoplasma-free.
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- 3-7 x 106 Raji-hEGFR cells in a cryovial or shipping flask.
- 1 ml of Blasticidin (10 mg/ml). Store at 4 °C or at -20 °C.
- 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi. Store at -20 °C.
IMPORTANT: If cells are shipped frozen (i.e. in a cryovial) and are not frozen upon arrival, contact InvivoGen immediately.
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