Practical in vitro sialylation of recombinant glycoproteins
AssigneeNeose Technologies, Inc.
InventorsJames C. Paulson, Robert J. Bayer, Eric Sjoberg
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Abstract
This invention provides methods for practical in vitro sialylation of glycoproteins, including recombinantly produced glycoproteins. The methods are useful for large-scale modification of sialylation patterns.
Claims58
A method of sialylating a saccharide group on a recombinant glycoprotein, the method comprising contacting a saccharide group which comprises a galactose or N-acetylgalactosamine acceptor moiety on a recombinant glycoprotein with a sialic acid donor moiety and a recombinant sialyltransferase in a reaction mixture which provides reactants required for sialyltransferase activity for a sufficient time and under appropriate conditions to transfer sialic acid from said sialic acid donor moiety to said saccharide group.
The method of claim, 13, wherein the bacterial sialyltransferase is a Neisseria meningitidis 2,3-sialyltransferase.
A method of sialylating a saccharide group on a recombinant glycoprotein, the method comprising contacting a saccharide group which comprises a galactose or an N-acetylgalactosamine acceptor moiety on a recombinant glycoprotein with a sialic acid donor moiety and a bacterial sialyltransferase in a reaction mixture which provides reactants required for sialyltransferase activity for a sufficient time and under appropriate conditions to transfer sialic acid from said sialic acid donor moiety to said saccharide group.
A method for in vitro sialylation of saccharide groups present on a glycoprotein, said method comprising contacting said saccharide groups with a sialyltransferase, a sialic acid donor moiety, and other reactants required for sialyltransferase activity for a sufficient time and under appropriate conditions to transfer sialic acid from said sialic acid donor moiety to said saccharide group, wherein said sialyltransferase is present at a concentration about 50 mU per mg of glycoprotein or less.
A method for in vitro sialylation of saccharide groups present on a glycoprotein, the method comprising contacting the saccharide groups with an ST3Gal III sialyltransferase, a sialic acid donor moiety, and other reactants required for sialyltransferase activity for a sufficient time and under conditions to transfer sialic acid from said sialic acid donor moiety to said saccharide group, wherein said ST3Gal III sialyltransferase is present at a concentration of less than about 50 mU per mg of glycoprotein.
Dependencies are inferred from the claim text.