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Cited by 88Hemophilia Treatment and ResearchViral Infectious Diseases and Gene Expression in InsectsProtein purification and stability

The first recombinant human coagulation factor VIII of human origin: human cell line and manufacturing characteristics

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Casademunt, Elisabeth, Martinelle, Kristina, Jernberg, Mats, Winge, Stefan, Tiemeyer, Maya, Biesert, Lothar, Knaub, Sigurd, Walter, Olaf +1 more·2012·European Journal of Haematology
DOI 10.1111/j.1600-0609.2012.01804.xPMC PMC3443369
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Abstract

Introduction: Since the early 1990s, recombinant human clotting factor VIII (rhFVIII) produced in hamster cells has been available for haemophilia A treatment. However, the post‐translational modifications of these proteins are not identical to those of native human FVIII, which may lead to immunogenic reactions and the development of inhibitors against rhFVIII. For the first time, rhFVIII produced in a human host cell line is available. Aim: We describe here the establishment of the first human production cell line for rhFVIII and the manufacturing process of this novel product. Methods and results: A human cell line expressing rhFVIII was derived from human embryonic kidney (HEK) 293 F cells transfected with an FVIII expression plasmid. No virus or virus‐like particles could be detected following extensive testing. The stringently controlled production process is completely free from added materials of animal or human origin. Multistep purification employing a combination of filtration and chromatography steps ensures the efficient removal of impurities. Solvent/detergent treatment and a 20 nm pore size nanofiltration step, used for the first time in rhFVIII manufacturing, efficiently eliminate any hypothetically present viruses. In contrast to hamster cell‐derived products, this rhFVIII product does not contain hamster‐like epitopes, which might be expected to be immunogenic. Conclusions: HEK 293 F cells, whose parental cell line HEK 293 has been used by researchers for decades, are a suitable production cell line for rhFVIII and will help avoid immunogenic epitopes. A modern manufacturing process has been developed to ensure the highest level of purity and pathogen safety.

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  • Human-cl rhFVIII produced in HEK293-F has a similar glycosylation profile to plasma-derived factor VIII and is deprived of α-gal and Neu5Gc.

    “It is produced in HEK293-F cell line and has shown similar glycosylation profile to the plasma-derived factor VIII, deprived of α-gal and Neu5Gc (Casademunt et al., 2012;Kannicht et al., 2013).”

    From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Casademunt et al., 2012

    The Methods and results passage identifies HEK 293 F cells as the source of the rhFVIII, and the Discussion says the product lacks Neu5Gc-type sialic acids and Galα1,3Gal (α-gal). Our copy does not establish that its glycosylation profile is similar to plasma-derived factor VIII; its discussion of an authentic human pattern of post-translational modifications does not make that comparison. No page numbers are available in the indexed passages. (LLM, conf=0.88)

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Authors
Casademunt, E., Martinelle, K., Jernberg, M., Winge, S., Tiemeyer, M., Biesert, L., Knaub, S., Walter, O., & Schröder, C.
Affiliations
  • Octapharma Munich Germany
  • Octapharma Stockholm Sweden
  • Octapharma Frankfurt Germany
  • Octapharma (Switzerland) (CH)
Corresponding author
Schröder, Carola
ORCID
Walter, Olaf
Venue
European Journal of Haematology
Publisher
Wiley
Published
2012-06-15
Volume
89
Issue
2
Pages
165-176
ISSN
  • 0902-4441 (ISSN-L)
  • 1600-0609
DOI
10.1111/j.1600-0609.2012.01804.x
PMC
PMC3443369
OpenAlex
W2091207267
Cited by
88
References
2
Citations by year
2026: 2 · 2025: 5 · 2024: 4 · 2023: 7 · 2021: 5 · 2020: 6 · 2019: 10 · 2018: 9 · 2017: 13 · 2016: 5 · 2015: 10 · 2014: 5 · 2013: 5 · 2012: 2
FWCI
4.69
Type
journal article
Language
en
Open access
Licence
onlinelibrary.wiley.com/termsAndConditions#vor (vor)
Locations
  • European Journal Of Haematology (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Health Sciences › Medicine › Hematology › Hemophilia Treatment and Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
Keywords
hemophilia A, human cell lines, protein expression, nanofiltration, virus removal, post-translational modification, immunogenicity
MeSH terms
Animals, Epitopes, Factor VIII, Cricetinae, Humans, Recombinant Proteins, Transfection, HEK293 Cells

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