Recombinant cell clone having increased stability in serum- and protein-free medium and a method of recovering the stable cell clone and the production of recombinant proteins by using a stable cell clone
AssigneeImmuno Aktiengesellschaft
InventorsManfred Reiter, Wolfgang Mundt, Friedrich Dorner
Life of this patent
Estimating the term…
Abstract
Disclosed are a stable recombinant cell clone which is 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.
Claims44
A recombinant CHO cell clone stable in serum- and protein-free medium for at least 40 generations and expressing a recombinant product.
A CHO cell culture comprising at least 90% of stable CHO recombinant cells, said stable recombinant cells being stable for more than 50 generations under serum- and protein-free conditions expressing recombinant product.
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.
A cell culture comprising more than 95% of stable recombinant cells, said stable recombinant cells being stable for more than 50 generations under serum- and protein-free conditions and expressing recombinant product.
A method for recovering a stable recombinant cell clone comprising the steps of
- multiplying a recombinant original cell clone in serum-containing medium so as to obtain a cell culture,
- culturing the cells of said cell culture under serum- and protein-free conditions,
- assaying said cell culture under serum- and protein-free conditions for producers of said recombinant product,
- cloning recombinant cell clones that are stable under serum and protein-free conditions, and
- multiplying said stable recombinant cell clones under serum- and protein-free conditions.
A method of recovering a stable recombinant cell clone comprising the steps of:
- multiplying a non-recombinant original cell under serum- and protein-free conditions,
- cloning a stable, non-recombinant cell clone under serum- and protein-free conditions,
- transfecting said stable cell clone with a recombinant nucleic acid and isolating stable transfectants,
- culturing said isolated stable transfectants in serum- and protein-free medium, and
- assaying said cultured transfectants for production stability.
A serum- and protein-free synthetic medium comprised of minimum medium, soybean peptone, glutamine, sodium hydrogencarbonate, ascorbic acid, ethanol amine and sodium selenite.
A cell culture in a serum- and protein-free synthetic medium, said serum- and protein-free synthetic medium being comprised of minimum medium, soybean peptone, glutamine, sodium hydrogencarbonate, ascorbic acid, ethanol amine and sodium selenite.
A method of producing a recombinant polypeptide in a cell culture comprising the steps of
- multiplying a stable starting cell clone expressing a recombinant polypeptide serum- and protein-free 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 CHO cell clone which has been cultivated in serum- and protein-free medium for at least 40 generations and which expresses a recombinant product.
Dependencies are inferred from the claim text.