Gal α1-3gal-containing N-glycans in glycoprotein products derived from CHO cells
First Claim
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1. A method for screening Chinese Hamster Ovary (CHO) cells for the ability to produce a target recombinant glycoprotein comprising glycans containing a target level of terminal galactose-alpha-1-3-galactose epitopes, the method comprising:
- (a) producing a target recombinant glycoprotein comprising one or more glycans by culturing CHO cells under conditions suitable for expression of the target recombinant glycoprotein by the CHO cells, wherein the CHO cells have not been genetically engineered to produce terminal alpha-galactosyl residues on glycans;
(b) treating the one or more glycans of the target recombinant glycoprotein with one or more exoglycosidases;
(c) detecting digested terminal galactose-alpha-1-3-galactose residues to thereby measure glycans containing terminal galactose-alpha-1-3-galactose residues produced by the CHO cells, and(d) selecting the CHO cells if a target level of terminal galactose-alpha-1-3-galactose residues is measured.
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Abstract
The present invention provides methods of evaluating CHO cells.
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Citations
18 Claims
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1. A method for screening Chinese Hamster Ovary (CHO) cells for the ability to produce a target recombinant glycoprotein comprising glycans containing a target level of terminal galactose-alpha-1-3-galactose epitopes, the method comprising:
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(a) producing a target recombinant glycoprotein comprising one or more glycans by culturing CHO cells under conditions suitable for expression of the target recombinant glycoprotein by the CHO cells, wherein the CHO cells have not been genetically engineered to produce terminal alpha-galactosyl residues on glycans; (b) treating the one or more glycans of the target recombinant glycoprotein with one or more exoglycosidases; (c) detecting digested terminal galactose-alpha-1-3-galactose residues to thereby measure glycans containing terminal galactose-alpha-1-3-galactose residues produced by the CHO cells, and (d) selecting the CHO cells if a target level of terminal galactose-alpha-1-3-galactose residues is measured. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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