Anti-IL-6R (Tocilizumab biosimilar - IgG1NQ isotype)

Non-glycosylated human IgG1NQ monoclonal antibody (mAb) against human IL-6R

Tocilizumab blocks IL-6R signaling

InvivoGen provides Anti-hIL-6R-hIgG1NQ, a monoclonal antibody (mAb) featuring:

• the variable region of tocilizumab targeting the human interleukin 6 receptor (IL-6R)
• a mutated non-glycosylated (NQ) IgG1 constant region with low effector functions

In the constant region, the asparagine (N) glycosylation sites were replaced by glutamine (Q) residues, resulting in the production of a non-glycosylated antibody. Glycosylation of an antibody does not affect antigen binding but is essential for Fc receptor-mediated activity [1]. In non-glycosylated antibodies, the effector mechanisms mediated by the Fc receptor types (FcgRI, FcgRII, FcgRIII) and the C1q component of the complement system are severely impaired or switched off [2].

InvivoGen's Anti-hIL6R-hIgG1NQ was generated by recombinant DNA technology, produced in Chinese hamster ovary (CHO) cells, and purified by affinity chromatography. Each lot is functionally tested by flow cytometry and cellular assays (see figures).

Key features of the Anti-hIL6R-hIgG1NQ

• Clinically-relevant variable region targeting human IL-6R (Tocilizumab)
• Human IgG1NQ constant region for low effector functions
• Inhibition of IL-6-dependent signaling has been validated in HEK-Blue™ IL-6 cellular assays
• The absence of bacterial contamination has been confirmed
   

IL-6R background

Tocilizumab (TCZ) is a recombinant, humanized monoclonal antibody (mAb) directed against both soluble and membrane-bound human IL-6R. TCZ exhibits anti-inflammatory activity by inhibiting the binding of the pro-inflammatory cytokine, IL-6  to its receptor [3]. IL-6 exerts its biological effects through IL-6R, and the transmembrane protein gp130. Despite only a few cells expressing IL-6R on their surface, many cells respond to IL-6 due to the existence of soluble IL-6R [4]. TCZ can block both modes of signaling [3]. TCZ has been approved for the treatment of inflammatory diseases and conditions such as rheumatoid arthritis (RA) and cytokine release syndrome (CRS), a side effect of CAR-T therapy. Furthermore, it is under investigation for the treatment of the late-stage hyperinflammation observed in severe COVID‑19 patients [5].

References:

1. Arnold J. et al., 2007. The impact of glycosylation on the biological function and structure of human immunoglobulins. Annu Rev Immunol 25:21-50.
2. Jefferis R., 2009. Glycosylation as a strategy to improve antibody-based therapeutics. Nat Rev Drug Discov. 8(3):226-34.
3. Sheppard, M. et al. 2017. Tocilizumab (Actemra). Hum Vaccin Immunother 13, 1972- 1988.
4. Rose-John, S. 2012. IL-6 trans-signaling via the soluble IL-6 receptor: importance for the pro-inflammatory activities of IL-6. Int J Biol Sci 8, 1237-1247.
5. Zhang, S. et al. 2020. Rational Use of Tocilizumab in the Treatment of Novel Coronavirus Pneumonia. Clin Drug Investig.

Figures

For your research, InvivoGen provides an Anti-hIL6R-To isotype family. This collection consists of mAbs comprising the variable region of Tocilizumab and differing constant regions of both native and engineered human isotypes. The isotypes differ in their effector functions, such as antibody-dependent cell-mediated cytotoxicity (ADCC), antibody‑dependent cellular phagocytosis (ADCP), and complement-dependent cytotoxicity (CDC). The Anti‑hIL6R-To isotype family will help you determine which isotype is the most suitable for your application.

Dose-dependent inhibition of HEK-Blue™ IL-6 cellular response. Anti-hIL6R‑To‑hIgG1NQ (0 - 1 μg/ml) was incubated with HEK-Blue™ IL-6 cells for 3 hours. Following this, human (h)IL-6 (0.3 ng/ml) was added to the cells. After overnight incubation, STAT3‑dependent SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution, a SEAP detection reagent. Data are presented as percentage (%) of the maximum response.

Specifications

Specificity: Human Interleukin-6 receptor (hIL-6R)

Clonality: Monoclonal antibodies

Clone: Tocilizumab (Anti-hIL-6R-hIgG1, kappa)

Control: Human IgG1NQ

Source: CHO cells

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

Tested application: Functional assay

Purification: Purified by affinity chromatography with protein G

Quality control:

• The complete sequence of each antibody construct has been verified.
• Each antibody lot has been functionally validated using HEK-Blue™ IL-6 cellular assays
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cellular assays.

Contents

Anti-hIL6R-To-hIgG1NQ purified monoclonal antibody is provided azide-free and lyophilized. It is available in two quantities:

• hil6rto-mab12: 100 µg
• hil6rto-mab12-1: 3 x 100 µg (300 µg)

 The product is shipped at room temperature.

 Upon receipt, store at -20°C.

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