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


OBJECTIVE
We describe here the establishment of the first human production cell line for rhFVIII and the manufacturing process of this novel product.


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.
Octapharma, Munich, Germany.
Elisabeth
E
Casademunt
Kristina
K
Martinelle
Mats
M
Jernberg
Stefan
S
Winge
Maya
M
Tiemeyer
Lothar
L
Biesert
Sigurd
S
Knaub
Olaf
O
Walter
Carola
C
Schröder
D000818:Animals; D006224:Cricetinae; D000939:Epitopes / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D005169:Factor VIII / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification*; D057809:HEK293 Cells; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification*; D014162:Transfection
D000939:Epitopes; D011994:Recombinant Proteins; C078147:F8 protein, human; D005169:Factor VIII