Recombinant cell clones having increased stability and methods of making and using the same
AssigneeBaxter Aktiengesellschaft
InventorsManfred Reiter, Wolfgang Mundt, Friedrich Dorner
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Abstract
Disclosed are a stable recombinant cell clones which are stable in serum- and protein-free medium for at least 40 generations, a biomass obtained by multiplying the stable cell clone under serum- and protein-free culturing conditions, and a method of preparing recombinant proteins by means of the biomass. Furthermore, the invention relates to a method of recovering stable recombinant cell clones.
Claims10
A method of producing a recombinant product under serum- and protein-free conditions on a large technical scale comprising the steps of:
- providing an isolated, stable recombinant original cell clone, said original cell clone being stable in serum- and protein-free medium for at least 40 generations and expressing a recombinant product,
- multiplying said original stable cell clone in serum- and protein-free medium so as to obtain a cell culture,
- culturing said cell culture containing stable cells in a bioreactor, thereby obtaining said recombinant product, and
- harvesting said recombinant product from a supernatant of said cell culture, wherein said serum- and protein-free medium comprises purified, ultrafiltrated soybean peptone having a molecular weight of ≦1000 Dalton.
A method for preparing a recombinant factor VIII protein or polypeptide having factor VIII activity from a mammalian cell culture wherein said mammalian cells have been transformed to express recombinant factor VIII, comprising the steps of:
- multiplying a mammalian recombinant cell clone stable in serum- and protein-free medium, wherein purified, ultrafiltered soybean peptone is added to the medium, so as to obtain a cell culture,
- culturing said cell culture containing stable cells in a bioreactor so as to obtain a cell culture, and
- harvesting said recombinant polypeptide from a supernatant of said cell culture.
A recombinant mammalian cell clone stable in serum- and protein-free medium for at least 40 generations and expressing a recombinant product, wherein said recombinant product is FVIII or a polypeptide having FVIII activity.
A recombinant mammalian cell clone stable in serum- and protein-free medium for at least 40 generations and expressing a recombinant product, wherein said recombinant product is vWF or a polypeptide having vWF activity.
A recombinant mammalian cell clone stable in serum- and protein-free medium for at least 40 generations and expressing a recombinant product, wherein said recombinant product is FVIII or a polypeptide having FVIII activity or vWF or a polypeptide having vWF activity.
Dependencies are inferred from the claim text.