Anti-PD-1 (Pembrolizumab biosimilar - IgG1 isotype)

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

Human IgG1 monoclonal antibody (mAb) against human PD-1

Anti-hPD1-Pem-hIgG1 features the constant region of the human IgG1 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].

Human IgG1 is the most abundant immunoglobin present in serum and binds with high affinity to the Fc receptor on phagocytic cells. The human IgG1 isotype displays high antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC).

Anti-hPD1-Pem-hIgG1 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

Activation of Jurkat-Lucia™ TCR-hPD-1 cells.
Raji-APC-hPD-L1 and Jurkat-Lucia™ TCR-hPD-1 cells were incubated with gradient concentrations of AntihPD-1 hIgG1 (Ni: Nivolumab variable region; Pem: Pembrolizumab variable region) or Anti-hPD-L1 mAbs for 6 hours. NFAT activation, reflecting the disruption of PD-1/PD-L1 inhibitory interaction, was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™.
The fold increase over non induced cells (no mAbs) is shown.

Specifications

Specificity: Targets cells expressing human PD-1

Clonality: Monoclonal antibody

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

Isotype: Human IgG1, kappa

Control: Human IgG1, kappa

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

• hpd1pe-mab1: 100 µg
• hpd1pe-mab1-03: 3 x 100 µg

Product is shipped at room temperature.

 Upon receipt, store at -20°C.