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Thromb Res. 2013 Jan;131(1):78-88. doi: 10.1016/j.thromres.2012.09.011. Epub 2012 Oct 8.

Characterisation of the post-translational modifications of a novel, human cell line-derived recombinant human factor VIII.

Author information

1
Octapharma, Berlin, Molecular Biochemistry, Germany. christoph.kannicht@octapharma.com

Abstract

INTRODUCTION:

Host cell lines used for recombinant protein expression differ in their ability to perform post-translational modifications (PTMs). The currently available recombinant human FVIII (rhFVIII) products are produced in mammalian, non-human cell lines. For rhFVIII, glycosylation and sulfation are vital for functionality and von Willebrand factor (VWF)-binding affinity. Here we present the characterisation of the PTMs of a novel, human cell line-derived recombinant human FVIII (human-cl rhFVIII). rhFVIII expression in a human cell line avoids expression of undesirable mammalian glycoforms like Galα1-3Galβ1-GlcNAc-R (α-Gal) and N-glycolylneuraminic acid (Neu5Gc), which constitute epitopes antigenic to humans.

MATERIALS AND METHODS:

We describe sulfation analysis, glycan profiling and characterisation using liquid chromatography-mass spectrometry and high performance anion exchange chromatography with pulsed amperometric detection.

RESULTS AND CONCLUSIONS:

Human-cl rhFVIII is confirmed to be sulfated and glycosylated comparable to human plasma-derived FVIII. Most importantly, human-cl rhFVIII is devoid of the antigenic Neu5Gc or α-Gal epitopes observed in Chinese Hamster Ovary- and Baby Hamster Kidney-derived rFVIII products. Both the avoidance of non-human glycan structures and the achievement of complete sulfation are proposed to lower the intrinsic immunogenicity of human-cl rhFVIII compared with current rFVIII products.

PMID:
23058466
DOI:
10.1016/j.thromres.2012.09.011
[Indexed for MEDLINE]

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