Enbrel ® (Etanercept) is a receptor fusion protein used in the treatment of Rheumatoid Arthritis, Polyarticular Juvenile Idiopathic Arthritis, Psoriatic Arthritis, Ankylosing Spondylitis and Plaque Psoriasis.

Etanercept is a homodimer made of two chains of 75kDa each, binding to TNF-α. The protein structure is complicated by the high degree of N- and O-glycosylation. Also, a N- and C-terminal sequence heterogeneity adds to the complexity of the protein analysis.

As the extended patent expires end of 2012, already many companies are developing biosimilar versions of this drug.

We have long-standing experience in the analysis of different variants of Enbrel originators and biosimilars. Due to the complexity of the protein, such a detailed analysis of the primary structure as well as the glycosylation can only be acheived by a combination of mass spectrometry, bioinformatics, HPLC and electrophoretic methods.

To support the development of Enbrel biosimilars or biobetters, Protagen has set up an analysis program in accordance with ICH Q6B guidelines, including method setup, validation of protein-specific methods, and release testing. This package includes

Protein quantification and determination of the extinction coefficient

Characterization of the protein primary structure including

  • confirming the amino acid sequence,
  • analysis of N- and C-terminal sequence heterogeneity,
  • determination of N- and O-glycosylation sites
  • quantificaiton of protein modifications,
  • confirming the disulfide bridging.

Characterization of the glycosylation pattern

  • N- and O-glycan mapping by mass spectromtry
  • N- and O-gylcan mapping by HPLC
  • Quantification of sialic acid levels
  • Linkage analysis by GC-MS

Characterization of the physicochemical properties

  • Intact protein mass by mass spectrometry
  • Charge isoforms by IEF, cIEF or IEX
  • Aggregation by AUC or SEC-MALLS

Your contact for Etanercept at Protagen Protein Services GmbH

Martin-Blueggel 150x100px

Martin Blüggel

Chief Executive Officer (CEO)
Our business development team will be happy to assist you with your project.

Phone: +49 (0) 231 9742-6100
Email: salesproteinservices@ProtagenProteinServices.com