|Raji-Null Cells||Unit size||Cat. code||Docs||Qty||Price|
Human lymphoblast cells - ADCC CD19/CD20/Control Target Cells
3-7 x 10e6 cells
Control Human B-cell line
Raji-Null cells were developed from the human Raji cell line. Raji-Null cells constitutively express different antigens at their cell surface that can be targeted by specific antibodies, notably to induce antibody-dependent cellular cytotoxicity (ADCC).
- Human CD20 is highly expressed by Raji cells. It has no known natural ligand and is thought to act as calcium channel enabling optimal B-cell responses . Human CD20 is a clinically-relevant target in mAb-mediated therapy (rituximab, ofatumomab, obinutuzumab) for non Hodgkin's B cell lymphoma or chronic lymphocytic leukaemia .
- Human CD19 is highly expressed by Raji cells. It acts as a co-receptor decreasing the threshold for antigen B-cell receptor (BCR)-dependent stimulation . Human CD19 is a clinical target for anti-hCD19-CD3 bis-specific antibodies (blinatumomab) in non Hodgkin's B cell lymphoma .
- Human PD-L1 (programmed cell death ligand 1; also known as CD274 or B7-H1) is expressed at low levels by Raji cells. This transmembrane protein is expressed by hematopoietic and nonhematopoietic cells, and is induced by pro-inflammatory cytokines, such as in the tumor micro-environment . Human PD-L1 is a clinically-relevant target in mAb-mediated therapy (atezolizumab, durvalumab, avelumab) for a wide range of cancers .
Features of Raji-Null cells:
- Constitutive expression of CD19 and CD20 at the cell surface
- Constitutive low expression of PD-L1 at the cell surface
- Stable expression of a resistance gene to Blasticidin
Applications for Raji-Null cells:
- ADCC target cells using anti-hCD19, anti-hCD20, anti-hPD-L1 Abs
- Negative control target cells in ADCC assays using monoclonal Abs other than anti-hCD19, anti-hCD20, anti-hPD-L1 Abs
- Target cells for anti-hCD19-CD3 bispecific Abs
Validation of Raji-Null cells:
- Functionally tested as target cells in ADCC assays using anti-human CD20 monoclonal Abs and Jurkat-Lucia™ NFAT-CD16 cells
- Functionally tested as negative control target cells in ADCC assays using monoclonal Abs other than anti-hCD20 (e.g. anti-hCTLA-4 or anti-hPD-1 Abs)
- Functionally tested as target cells in T-cell activation assay using anti-hCD19-CD3 bi-specific Ab and Jurkat-Lucia™ NFAT cells.
1. Bae J. et al. 2005. Identification of CD19 and CD20 peptides for induction of antigen-specific CTLs against B-cell malignancies. Clin. Cancer. Res. 11:1629-38.
2. Almagro J.C. et al. 2018. Progress and challenges in the development of antibodies for cancer therapy. Front. Immunol. 8:1751.
3. Bargou R. et al. 2008. Tumor regression in cancer patients by very low doses of a T cell-engaging antibody. Science. 321(5891):974-7.
4. Ribas A. and Wolchock J.D., 2018. Cancer immunotherapy using checkpoint blockade. Science. 359:1350-55.
Comparison of ADCC potency for native and engineered anti-human CD20 antibody isotypes: Raji-Null cells were incubated with gradient concentrations of Anti-hCD20 or Anti-β-galactosidase (β-gal) mAbs for 1 hour. Jurkat-Lucia™ NFAT-CD16 effector cells were then co-incubated with targets 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
Figure 2. Comparison of ADCC potency of anti-human CTLA-4, PD-1, or PD-L1 IgG1 antibodies on Raji-derived target cells: Raji-hCTLA4, Raji-hPD-1, Raji-hPD-L1 or Raji-Null control cells were incubated with gradient concentrations of anti-hCTLA4-IgG1, anti-hPD-1-IgG1, or anti-hPD-L1-IgG1 mAbs for 1 hour. Jurkat-Lucia™ NFAT-CD16 effector cells were then co-incubated with targets 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 are shown, and responses with Raji-Null cells are normalized on appropriate antigen-expressing cells.
- Human CD20 expression has been verified by flow-cytometry.
- Induction of antibody-dependent cellular cytotoxicity (ADCC) has been validated using InvivoGen’s anti-hCD20-hIgG1 antibody and Jurkat-NFAT Lucia™ CD16 reporter cell line.
- The stability for 20 passages following thawing has been verified.
- Raji-Null cells are guaranteed mycoplasma-free.
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- 1 vial of Raji-Null cells (3-7 x 106 cells) in Freezing Medium
- 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: Cells are shipped frozen. If cells are not frozen upon arrival, contact InvivoGen immediately.
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