HER2-expressing Raji Cells

HER2-expressing B cells

Raji-HER2 cells were developed from the human Raji cell line, a human B lymphocyte-derived cell line, and engineered to stably overexpress the human HER2 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.

Human epidermal growth factor receptor 2 (HER2), also known as ERBB2 or neu, is a member of the EGFR/ERBB family of receptor tyrosine kinases. Due to its overexpression on the surface of not only breast cancer cells but a variety of solid tumors, HER2 is both an oncogene and a tumor-associated antigen [1, 2]. Heterodimerization of HER2 with other members of the EGFR family results in the activation of a variety of potent proliferative and anti-apoptotic signaling pathways. Therefore, HER2 is a major driver of tumor development and progression [1]. There is a variety of approved monoclonal antibody (mAb) therapies (e.g. Trastuzumab) successfully used in the treatment of HER2-positive cancers (e.g. breast and gastric) [2]. Thus, HER2 remains an important therapeutic target for the development of more potent mAbs, specifically with optimized effector functions for improved tumor-directed ADCC [2].

Features of Raji-HER2 cells:

Surface expressed markers in Raji-HER2 cells

• Stable overexpression of the human HER2 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-HER2 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-HER2 cells:

• Overexpression of HER2 verified by flow cytometry
• Functionally tested as target cells in ADCC assays using anti-human HER2 mAbs and Jurkat-Lucia™ NFAT-CD16 cells
• Guaranteed mycoplasma-free

​References:

1. Gutierrez, C. & Schiff, R. 2011. HER2: biology, detection, and clinical implications. Arch Pathol Lab Med 135, 55-62.
2. Oh, D.Y. & Bang, Y.J. 2020. HER2-targeted therapies - a role beyond breast cancer. Nat Rev Clin Oncol 17, 33-48.

Figures

Validation of the expression of HER2 by Raji-HER2 cells. Raji-Null (A) and Raji‑HER2 (B) cells were incubated with a PE-conjugated Anti-HER2 mAb for 30 minutes. The binding affinity was then measured using flow cytometry.

Comparison of ADCC potency for native and engineered anti-HER2 antibody isotypes. Raji-HER2 cells were incubated with gradient concentrations of Anti-HER2-Tra mAbs, which feature the Trastuzumab variable region, or an Anti-β-galactosidase (β-Gal) mAb 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), 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:

• HER2 expression has been verified by flow cytometry.
• Induction of antibody-dependent cellular cytotoxicity (ADCC) has been validated using anti-HER2 antibodies and InvivoGen's Jurkat-NFAT Lucia™ CD16 reporter cell line.
• The stability for 20 passages following thawing has been verified.
• Raji-HER2 cells are guaranteed mycoplasma-free.

Contents

• 3-7 x 10⁶ Raji-HER2 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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