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US 10,260,056 B2Granted
Cleavage of fucose in N-glycans
AssigneeNew England Biolabs, Inc.
InventorsChristopher H. Taron, Saulius Vainauskas, Xiaofeng Shi
Life of this patent
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Abstract
Provided herein is an α-fucosidase that can cleave a conjugate comprising an N-glycan and a label where the label is added by amine reactive chemistry. The α-fucosidase also has an accelerated reaction time using Schiff base labeled N-glycans compared with bovine kidney fucosidase. A reaction mix, enzyme mix and kit comprising the α-fucosidase are provided, as well as a method for analyzing glycoproteins. The α-fucosidase finds particular use in analyzing the N-glycans of therapeutic glycoproteins.
Claims17
A reaction mix comprising:
- (a) an α-L-fucosidase comprising an amino acid sequence that is identical to amino acids 23-359 of the sequence of SEQ ID NO:1;
- and
- (b) a conjugate comprising an N-glycan and a label that are joined by an carbamide bond.
An enzyme mix comprising:
- (a) an α-fucosidase comprising an amino acid sequence that is identical to amino acids 23-359 of the sequence of SEQ ID NO:1;
- and
- (b) one or more exoglycosidases that are not encoded by the genome of Omnitrophica, selected from the group consisting of:
- α2-3 neuraminidase S;
- α2-3,6,8,9 neuraminidase A;
- α1-3,4,6 galactosidase;
- β1-4 galactosidase;
- β-N-acetylglucosaminidase S;
- and
- α1-2,3,6 mannosidase.
7 more elements
A kit, comprising:
- a. an α-fucosidase comprising an amino acid sequence that is identical to amino acids 23-269 of the sequence of SEQ ID NO:1;
- and
- b. a label comprising a reactive carbamate.
Dependencies are inferred from the claim text.