Anti-EGFR (Cetuximab biosimilar - IgG1 isotype)

Human IgG1 monoclonal antibody (mAb) against human EGFR

Anti-hEGFR-hIgG1 is the biosimilar of the clinical antibody cetuximab. Anti-hEGFR-hIgG1 features the constant region of the human IgG1 isotype and the variable region of cetuximab. Cetuximab is a chimeric human/mouse IgG1 monoclonal antibody that targets epidermal growth factor receptor (EGFR), a cell surface receptor overexpressed in many types of cancer.

EGFR is activated by binding specific ligands, including epidermal growth factor and transforming growth factor-α. Activation of EGFR promotes cell proliferation and survival, as well as angiogenesis, leading to tumor growth and metastasis.

The binding of cetuximab to EGFR blocks ligand/receptor binding and induces receptor internalization and subsequent degradation.

Consequently, cetuximab blocks downstream pathways which regulate cell growth and angiogenesis. In addition, cetuximab induces cell death through antibody-dependent cell-mediated cytotoxicity (ADCC) [1, 2]. Cetuximab has been approved by the FDA for the treatment of metastatic colorectal cancer and metastatic squamous cell carcinoma of the head and neck [3].

Anti-hEGFR-hIgG1 was generated by recombinant DNA technology. It has been produced in Chinese hamster ovary (CHO) cells and purified by affinity chromatography with protein G.

More isotypes of this antibody are available and can be used for comparison of biological activities such as ADCC (see in the 'upon request' section).

References:

Kurai J. et al., 2007. Antibody-dependent cellular cytotoxicity mediated by cetuximab against lung cancer cell lines. Clin Cancer Res. 3(5):1552-61.
Kimura H. et al., 2007. Antibody-dependent cellular cytotoxicity of cetuximab against tumor cells with wild-type or mutant epidermal growth factor receptor. Cancer Sci. 98(8):1275-80.
Vincenzi B. et al., 2010. Cetuximab: from bench to bedside. Curr Cancer Drug Targets. 10(1):80-95.

Figures

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

Specificity: Targets cells expressing human EGFR

Clonality: Monoclonal antibody

Clone: Cetuximab (Anti-hEGFR-hIgG1, kappa)

Isotype: Human IgG1, kappa

Control: Human IgG1

Source: CHO cells

Formulation: 0.2 µm filtered solution in 68 mM sodium phosphate buffer (pH 7.4) with 91 mM glycine, 5% w/v saccharose, and stabilizing agents

Purity: Purified by affinity chromatography with protein G

Tested applications: Flow cytometry and ADCC

Quality control:

• Binding of Anti-hEGFR-hIgG1 to human EGFR has been tested using flow cytometry.
• The complete sequence of this antibody has been verified.
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.

Contents

Anti-hEGFR-hIgG1 purified monoclonal antibody is provided azide-free and lyophilized. It is available in two quantities:

• hegfr-mab1: 100 µg
• hegfr-mab1-1: 1 mg

Product is shipped at room temperature

 Upon receipt, store at -20°C.

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