Anti-PD-1 (Pembrolizumab biosimilar - IgG4 (S228P) isotype)

Human IgG4 (S228P) engineered monoclonal antibody (mAb) against human PD-1

Principle of PD-1/PD-L1 cellular assay
(click to enlarge)

Anti-hPD1-Pem-hIgG4 (S228P) is the biosimilar of the clinical antibody pembrolizumab. Anti-hPD1-Pem-hIgG4 (S228P) features the constant region of the human IgG4 (S228P) isotype and the variable region of pembrolizumab. Pembrolizumab is a humanized IgG4 monoclonal antibody that contains an engineered hinge region mutation (S228P) designed to prevent the exchange of IgG4 molecules. This antibody targets the PD-1 receptor found on activated T cells, B cells, and myeloid cells. Under normal physiological conditions, PD-1 negatively regulates T cell activation thereby preventing autoimmunity [1].

Under pathological conditions, cancer cells produce PD-L1 (programmed cell death 1 ligand 1), the agonist that binds and activates PD-1. Activated PD-1 enables the cancer cells to evade the immune system. Pembrolizumab binds and blocks the activation of the PD-1 receptor, thereby resulting in the activation of T cells [2]. Pembrolizumab has been approved by the FDA for the treatment of metastatic malignant melanoma and is currently under regulatory review in the EU [3].

Anti-hPD1-Pem-hIgG4 (S228P) was generated by recombinant DNA technology. It has been produced in Chinese hamster ovary (CHO) cells and purified by affinity chromatography. More isotypes are available and can be used for comparison of biological activities such as ADCC (see below or in the 'upon request' section).

References:

1. McDermott D. & Atkins M. 2013. PD-1 as a potential target in cancer therapy. Cancer Med. 2(5): 662–673.
2. Tumeh PC. et al., 2014. PD-1 blockade induces responses by inhibiting adaptive immune resistance. Nature. 515(7528):568-71.
3. Poole RM., 2014. Pembrolizumab: first global approval. Drugs. 4(16):1973-81.

Figures

Binding of  Anti-hPD1-Pem-hIgG4 (S228P) mAb to hPD-1. EL4-hPD-1 cells were incubated for 30 minutes at 4°C with 125 ng of Anti-hPD1-Pem-hIgG4 (S228P) or an isotype control. Subsequently, a secondary PE-labeled antibody was added and incubated at 4°C for 30 minutes. The binding affinity was assessed using flow cytometry.

Specifications

Specificity: Targets cells expressing human PD-1

Clonality: Monoclonal antibody

Clone: Pembrolizumab (Anti-hPD1-hIgG4 (S228P), kappa)

Isotype: Human IgG4 (S228P), kappa

Control: Human IgG4 (S228P)

Source: CHO cells

Formulation: 0.2 μm filtered solution in a sodium phosphate buffer with glycine, saccharose, and stabilizing agents.

Purity: Purified by affinity chromatography with protein G

Applications: Anti-hPD1-Pem-hIgG4 (S228P) binds and blocks ligand activation of the PD-1 receptor on the surface of activated T cells.

Quality control:

• The recognition of PD-1 with this antibody 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-hPD1-Pem-hIgG4 (S228P) purified monoclonal antibody is provided azide-free and lyophilized. It is available in two quantities:

• hpd1pe-mab14: 100 µg
• hpd1pe-mab14-1: 1 mg

 The product is shipped at room temperature.

 Upon receipt, store at -20°C.

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