Anti-hIL6R (Tocilizumab) isotype family

Recombinant anti-hIL-6R antibody (Tocilizumab)

Tocilizumab blocks IL-6R signaling

InvivoGen provides a family of Anti-hIL6R-derived monoclonal antibodies (mAbs) in multiple human isotypes. These mAbs feature:

• the variable region of tocilizumab, which specifically targets the human interleukin-6 receptor (hIL6R)
• a constant region that mediates different effector functions depending on the isotype: hIgG1, hIgG1NQ or IgA2

InvivoGen's hIL6R mAbs have been generated by recombinant DNA technology, produced in CHO cells, and purified by affinity chromatography.

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 [1]. 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 [2]. TCZ can block both modes of signaling [1]. 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 [3].

Key features of the Anti-hIL6R isotype family:

• Clinically-relevant variable region targeting human IL-6R (Tocilizumab).
• Inhibition of IL-6-dependent signaling by Anti-hIL6R-To mAbs has been validated in HEK-Blue™ IL-6 cellular assays.
• The absence of bacterial contamination has been confirmed.

InvivoGen’s products are for research use only, and not for clinical or veterinary use.

Read our reviews on COVID-19 and learn more about SARS-CoV-2 infection cycle, immune responses, and potential therapeutics.

References:

1. Sheppard, M. et al. 2017. Tocilizumab (Actemra). Hum Vaccin Immunother 13, 1972- 1988.
2. 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.
3. 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-hIgG1 (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.

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.

Dose-dependent inhibition of HEK-Blue™ IL-6 cellular response. Anti-hIL6R‑To‑hIgA2 (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
Source: CHO cells
Formulation: 0.2 μm filtered solution in a sodium phosphate buffer with glycine, saccharose, and stabilizing agents.
Purification: 

• hIgG1: purified by affinity chromatography with protein G
• hIgG1NQ: purified by affinity chromatography with protein G
• hIgA2: purified by affinity chromatography with peptide M

Quality control:

• The complete sequence of each antibody construct has been verified.
• Each antibody 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

• 100 µg purified antibody, azide-free, and lyophilized

 The product is shipped at room temperature.

 Store lyophilized antibody at -20 °C.

 Lyophilized product is stable for at least 1 year.