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US 9,273,292 B2Granted
Chinese hamster ovary cell lines
AssigneeAgency for Science, Technology and Research (A*Star)
InventorZhiwei Song
Life of this patent
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Abstract
We provide a Chinese Hamster Ovary (CHO) cell which is capable of higher protein sialylation compared to a wild type Chinese Hamster Ovary cell, such as in the presence of functional GnT 1, in which the CHO cell is obtainable by selection with Ricinus communis agglutinin I (RCA-I).
Claims12
A CHO cell comprising a mutation in the GnT I gene selected from the following mutations at the specified position of a GnT I nucleic acid sequence of GenBank Accession Number:
- AF343963:
- (a) a C to T transition at nucleic acid sequence position 1015;
- (b) a G to C transversion at nucleic acid sequence position 1300;
- (c) an A to C transversion at nucleic acid sequence position 638;
- (d) a C to G transversion at nucleic acid sequence position 784;
- (e) a T to A transversion at nucleic acid sequence position 811;
- (f) an insertion at nucleic acid sequence position 706 resulting in a frame shift from nucleic acid sequence position 236 of the encoded amino acid sequence;
- (g) a G to A transition at nucleic acid sequence position 246;
- (h) a G to A transition at nucleic acid sequence position 258;
- or (i) an A to T transversion at nucleic acid sequence position 859, in which the CHO cell further comprises a nucleic acid encoding a protein of interest comprising a glycoprotein in an expression vector and a nucleic acid encoding a functional GnT I sequence.
7 more elements
A method of providing a CHO cell or cell line comprising a mutation in the GnT I gene (GenBank accession number:
- AF343963), the method comprising culturing CHO cells in the presence of Ricinus communis agglutinin I (RCA-I), in which a recombinant protein of interest expressed by the resulting CHO cell or cell line in the presence of an introduced nucleic acid encoding functional GnT 1 comprises a higher sialylation than a protein of interest expressed by a wild-type Chinese Hamster Ovary cell.
Dependencies are inferred from the claim text.