Human EGFR-expressing Raji 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 [1]. 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:

Surface expressed markers in 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

​References:

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.

Figures

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.

Specifications

Antibiotic resistance: Blasticidin

Growth medium: IMDM, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum (FBS; 30 min at 56 °C), 10 μg/ml Blasticidin, Pen-Strep (100 U/ml-100 µg/ml), 100 µg/ml Normocin™

Test medium: IMDM, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated FBS, Pen-Strep (100 U/ml-100 µg/ml)

Quality control:

• 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.

Contents

• 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.

Shipped on dry ice (Europe, USA & Canada)

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