Methods for selecting eukaryotic cells expressing a heterologous protein
AssigneeNovartis AG
InventorsThomas Jostock, Hans-Peter Knopf
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Abstract
The present invention pertains to a method for selecting at least one eukaryotic host cell expressing a product of interest. The method entails providing a plurality of host cells which are dependent upon folate uptake and contain polynucleotides encoding for the product of interest and a DHFR enzyme. The cells are cultured in a selective culture medium containing at least an inhibitor of DHFR and folate. The expressed cells are then selected.
Claims15
A method for selecting at least one eukaryotic host cell expressing a product of interest, comprising at least the following steps:
- (a) providing a plurality of eukaryotic host cells, wherein cellular viability of said host cells is dependent upon folate uptake, wherein said eukaryotic host cells comprise at least
- (i) an introduced polynucleotide encoding a product of interest and
- (ii) an introduced polynucleotide encoding a DHFR enzyme;
- (b) culturing said plurality of eukaryotic host cells in a selective culture medium comprising at least an inhibitor of DHFR and folate in a limiting concentration;
- (c) selecting at least one eukaryotic host cell expressing the product of interest.
The method according to claim lwherein said eukaryotic host cell is a CHO host cell.
Dependencies are inferred from the claim text.