Enbrel (Etanercept) Biosimilars

Enbrel Biosimilars

Enbrel (Etanercept) is a complex therapeutic fusion protein consisting of the Fc region of the human IgG1 antibody fused to a portion of the tumor necrosis factor receptor (TNFR). Its therapeutic activity comes from its ability to bind and inhibit as a tumor necrosis factor alpha (TNFα). TNFα is a potent pro-inflammatory cytokine involved in a number of inflammatory diseases. Several Enbrel biosimilars have been approved by the EMA and FDA.

The manufacture of more affordable Enbrel biosimilars offers more patients the opportunity to be treated with the advanced biopharmaceuticals. However, biosimilar development and characterization is complex and time-consuming. The production of Enbrel biosimilars presents additional hurdles, as it can be challenging to achieve robust expression of fusion proteins. Therefore, a reliable cell line development provider is key to the successful production of Enbrel biosimilars.

Sartorius has extensive experience in providing support throughout all stages of development and manufacture of TNFα-targeting biosimilars. Our CellcaCHO expression platform has demonstrated success in delivering high-yielding cell lines, helping you optimize downstream process development. We can also facilitate the seamless transition into our GMP cell banking and characterization services accelerates your Enbrel biosimilar program towards your first GMP batch.


Simplify Your Biosimilar Characterization

Our panel of ready-to-use, pre-qualified assays facilitate comprehensive characterization of Enbrel biosimilars 

  • Uncover valuable insights into your biosimilar 

  • Accelerate development times 

  • Meet regulatory requirements 

  • Boost confidence in your product 

  • Limit the risks associated with biosimilar development

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Characteristics of Enbrel

TNFα is a small signaling protein with a significant role in normal immune function. Elevated levels of TNF are found in involved tissues and fluids of patients with some disorders associated with chronic inflammation.  

Enbrel acts by inhibiting TNF-α, and is permitted for the treatment of several chronic inflammatory conditions including severe rheumatoid arthritis, juvenile idiopathic arthritis, severe plaque psoriasis, and ankylosing spondylitis. 

Enbrel is a fusion protein consisting of the ligand-binding portion of the 75-kilodalton TNFR (TNFR2/p75) genetically fused to the Fc portion of human IgG1. It primarily works by competitively binding TNFα to inhibit its actions. The TNFR component binds with high affinity to the soluble form of TNFα, acting as a decoy receptor and preventing TNFα from binding to cellular TNFRs.   

TNFα is an exogenous cytokine that contributes to the regulation of the cells of the immune system. It is produced by several immune cells in response to infection and inflammation. Normally, when TNF binds its receptors on the surface of immune cells, it initiates a complex signaling network, including the activation of NFκB, MAPK, and apoptosis pathways.  

Ultimately, intracellular TNFα signaling leads to amplified inflammation. Enbrel sequesters TNFα, limiting its effects.

Sartorius can support your Enbrel biosimilar project from cell-line development through to complete analysis of your product. Our off-the-shelf characterization solutions were designed to meet regulatory requirements. 

  • Physicochemical and structural assays reveal detailed insights into the composition of your biosimilar   

  • Binding assays quantify and evaluate the binding of your biosimilar to TNFα and other components of the immune system. 

  • Bioassays measure the biological activity of your Enbrel biosimilar, giving you valuable data to support your comparability program. 

Assay Types

Physicochemical and structural characterization is a crucial step in determining the potential biological activity, stability, and safety of your biosimilar.   

Our versatile platform of physicochemical methods provides the basis for your comparability study.  With options suited for clone selection through to formal comparability, we can build detailed insights into the properties of your biosimilar.    

The combination of our off-the-shelf physicochemical and structural analyses and our binding and bioassays allows you to evaluate the structure-function relationship and ticks the regulators' box for orthogonal comparability.   

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We employ range of cell-based neutralization assays which offer detailed characterization of Embrel’s TNF inhibition and its downstream effects. These include: 

  • Measurement of cell death/viability using L929 cells 

  • Measurement of cell death/viability using U937 cells 

  • Measurement of apoptosis using U937 cells 

  • Measurement of ELAM-1 expression (marker of cell adhesion) using HUVEC cells 

  • Measurement of ICAM-1 (marker of cell adhesion) Expression) 

  • Measurement of VCAM-1 (marker of cell adhesion)  

We also offer both Complement Dependent Cytotoxicity (CDC) and Antibody Dependent Cellular Cytoxicity (ADCC) testing to enhance your comparability studies. These assays use engineered cell lines expressing membrane-bound TNFα as the target cells.  

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Binding assays are employed to study a number of different features of Enbrel and play a vital role in characterization and comparability studies.  Sartorius offers a comprehensive range of off-the-shelf binding methods to measure the binding of Enbrel to TNF-Alpha as well as C1q, the Fc Gamma Receptors (FcR) and the neonatal Fc Receptor (FcRn). 

  • Enbrel TNF-Alpha Binding Assays - Our Enbrel TNFα binding assays use an upgraded version of the traditional ELISA on the Mesoscale Discovery (MSD) platform to report the relative binding of your biosimilar compared to the innovator Enbrel material as a percentage of a designated reference lot, with comprehensive parallelism assessments. This method can be adapted for use as a traditional ELISA platform to support lot release at contract manufacturing facilities, where required. The assay can be performed using recombinant TNFα or cells engineered to express membrane-boundTNFα. 

  • Enbrel Fc-Gamma Receptor Assays 

The Enbrel Fc-Gamma Receptor assays offered by BioOutsource are performed using SPR instruments, which are versatile, label-free systems with exceptional sensitivity. The following Enbrel (etanercept) Fc-Gamma Receptor assays are offered by BioOutsource: 

  • Enbrel Fc-Gamma Receptor I (FcRI) binding assay by SPR 

  • Enbrel Fc-Gamma Receptor IIa (FcRIIa) binding assay by SPR 

  • Enbrel Fc-Gamma Receptor IIb (FcRIIb) binding assay by SPR 

  • Enbrel Fc-Gamma Receptor IIIa (V) (FcRIIIa V) binding assay by SPR 

  • Enbrel Fc-Gamma Receptor IIIa (F) (FcRIIIa F) binding assay by SPR 

  • Enbrel Fc-Gamma Receptor IIIb (FcRIIIb) binding assay by SPR 

  • Enbrel FcRn Assays 

We have extensive experience analyzing Enbrel biosimilar and innovator material binding to FcRn by SPR. Our Enbrel FcRn binding assay services can be used to determine the Binding Affinity Constant (KD). 

  • Enbrel C1q Assays 

Enbrel (etanercept) mediates its activity through the interaction with the TNF-Alpha and following binding to the membrane bound form, Enbrel is capable of activating the complement cascade and inducing cell lysis via complement dependent cytotoxicity. Activation of complement is experimentally determined using the C1q binding ELISA assay. 

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Related Assets


Physicochemical and Biological Analytical Comparability for Biosimilars

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