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CE-SDS analysis of Etanercept using BioPhase 8800 system

CE-SDS analysis of Etanercept using BioPhase 8800 system


Etanercept has been widely used in the treatment of plaque psoriasis, psoriatic arthritis, ankylosing spondylitis, and rheumatoid arthritis for over 20 years.1, 2 It works as a tumor necrosis factor (TNF) antagonist by binding to TNF molecules and preventing them from binding to the cell-bound receptors—which could otherwise lead to autoimmune disease when de-regulated TNF production occurs.3 Etanercept is a heavily glycosylated fusion protein consisting of the extracellular ligand-binding domain of human TNF receptor 2 (TNFR2) and the Fc domain of human IgG1.4 There are 2 N-glycosylation sites and 13 potential O-glycosylation sites on the TNFR2 domain and one N-glycosylation site on the Fc domain.4 The glycosylation alone represents approximately 30% of the total molecular weight of etanercept.5 The fusion protein is also a homodimer where disulfide bonds connect the two monomers. These properties combined make the protein structure very complex and highly heterogeneous. The physicochemical characterization of such molecules is very challenging.

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