BioPharmaSpec provides a complete solution for Etanercept/ Enbrel comparability, biosimilarity and characterization studies.
Etanercept (trade name Enbrel) is a recombinant protein made by fusing the TNF-receptor and the constant region of the IgG1 antibody. It is a large biopharmaceutical with a molecular weight of 150 kDa that, similar to Humira, Simponi and Remicade, inhibits tumor necrosis factor alpha (TNFα). TNFα is central to the inflammatory response and plays a major role in inflammatory/ autoimmune diseases such as arthritis, plaque psoriasis and ankylosing spondylitis. Etanercept is therefore used to treat certain inflammatory diseases.
Biosimilar Characterization Considerations
The guidelines state that Etanercept/ Enbrel comparability and characterization should include determination of physicochemical and structural properties, purity, impurities and quantity in line with ICH Topic Q6B:
- Primary (de novo) Amino Acid Sequence
- Amino Acid Composition
- Terminal Amino Acid Sequence
- Peptide Map
- Disulfide Bridges
- Carbohydrate Structure
- Secondary and Tertiary Structure
Functional Testing by Custom Biologics
- Caspase 3/Apoptosis Assay
- NFkB Reporter Assay
- TNFα Neutralization Assay
- Antibody-Dependent Cell Cytotoxicity (ADCC) Reporter Bioassay
- Complement-Dependent Cytotoxicity (CDC) Assay
- Proliferation Assays
- Binding Assays
Product Specific Technical Considerations
Particular attention should be paid to the Post Translational Modifications (PTMs) including:
- Disulfide bridges (particularly in the TNFα region)
- C-terminal Lysine
- Glycosylation (in particular, levels of galactosylation, fucosylation and sialylation, N-glycosylation site profiling and occupancy and O-glycosylation structure and site occupancy).