Anti-TNF-α (Adalimumab biosimilar - IgG1 isotype)

Human IgG1 monoclonal antibody (mAb) against human TNF-α

Neutralizing monoclonal antibody against human TNF-α

Anti-hTNF-α-hIgG1 is the biosimilar of the clinical antibody adalimumab. Anti-hTNF-α-hIgG1 a neutralizing monoclonal antibody (mAb) featuring the constant region of the human IgG1 (hIgG1) isotype and the variable region of adalimumab, a fully human therapeutic mAb that targets human tumor necrosis factor-alpha (hTNF-α). The variable region of adalimumab blocks the interaction of soluble and membrane-bound TNF-α with its receptors TFNR1 and TNFR2, thereby downregulating the inflammatory responses [1,2]. 

Anti-hTNF-α-hIgG1 was generated by recombinant DNA technology. It is produced in CHO cells and purified by affinity chromatography with protein G. The neutralizing activity of Anti-hTNF-α-hIgG1 was determined using recombinant human TNF-α and HEK-Blue™ TNF-α cells. TNF-α is a pleiotropic cytokine that notably induces NF-κB-mediated production of pro-inflammatory cytokines [1,2].

 More details

Human IgG1 Isotype:

The human IgG1 isotype displays a high binding affinity for the Fc receptor on phagocytic cells and is highly potent for Fc-mediated antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and complement-dependent cytotoxicity (CDC). 

 Learn more about antibody isotype effector functions by reading our flyer on Clinically relevant monoclonal antibodies.

References:

1. Steeland S., et al. 2018. A new venue of TNF targeting. Int. J. Mol. Sci. 19:1442.
2. Shealy D.J. & Visvanathan S. 2008. Anti-TNF antibodies: lessons from the past, roadmap for the future. Therapeutic Antibodies (book). 101-129.

Figures

Evaluation of hTNF-α inhibition. Increasing concentrations of Anti-hTNF-α-IgG1 were incubated for 30 minutes with the recombinant cytokine hTNF-α (30 pg/ml) before the addition of HEK-Blue™ TNF-α cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown in percentage (%) of activity.

Specifications

Specificity: Human TNF-α (hTNF-α)

Clonality: Monoclonal antibody

Isotype: Human IgG1, kappa

Control: Anti-β-Gal-hIgG1

Source: Chinese hamster ovary (CHO) cells

Purification: Affinity chromatography with protein G

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

Tested application: Blocking & Neutralization of hTNF-α signaling

Quality control:

• Human IgG1 isotype has been confirmed by ELISA.
• The neutralization of hTNF-α signaling pathway has been confirmed using cellular assays with the HEK-Blue™ TNF-α cells.
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.

Contents

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

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

 The product is shipped at room temperature.

Upon receipt, store lyophilized antibody at -20°C.

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Details

Tumor necrosis factor-alpha (TNF-α) is a pleiotropic cytokine involved in necrotic and apoptotic cell death, cellular differentiation, inflammation, and regulation of immune cell activity [1]. TNF-α is mainly produced by activated monocytes, macrophages, and T cells. It is first synthesized as a membrane-bound molecule that forms a compact homotrimer through non-covalent interactions. The trimeric membrane-bound form is cleaved by tumor necrosis factor-alpha converting enzyme (TACE) releasing the soluble trimer [2]. Both the membrane-bound and soluble TNF-α bind homotrimeric transmembrane receptors, TNFR1 or TNFR2, triggering signaling pathways that involve TRADD, TRAF2, and RIP, and leading to the activation of NF-κB and MAPK pathways. Importantly, membrane-bound TNF can also mediate a reverse "outside-to-inside" signaling, although this mechanism remains to be fully elucidated [2].

References:

1. Steeland S., et al. 2018. A new venue of TNF targeting. Int. J. Mol. Sci. 19:1442.
2. Shealy D.J. & Visvanathan S. 2008. Anti-TNF antibodies: lessons from the past, roadmap for the future. Therapeutic Antibodies (book). 101-129.