Site-specific glycoengineering of targeting moieties
AssigneeGenzyme Corporation
InventorsLuis Avila, Clark Pan, Huawei Qiu, Qun Zhou
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Abstract
The current disclosure provides binding polypeptides (e.g., antibodies), and targeting moiety conjugates thereof, comprising a site-specifically engineered glycan linkage within native or engineered glycans of the binding polypeptide. The current disclosure also provides nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided.
Claims27
A method of making a PEGylated binding polypeptide comprising at least one modified glycan, wherein the at least one modified glycan comprises at least one moiety of Formula (IV):
- the method comprising reacting an effector moiety of Formula (I):
- NH2-Q-CON-X Formula (I),
- with a precursor binding polypeptide comprising an oxidized glycan
- wherein:
- A) Q is NH or O;
- B) CON is a connector moiety;
- C) X is a moiety comprising a PEG moiety;
- D) Gal is a galactose moiety;
- and
- E) Sia is a sialic acid moiety,
- wherein the binding polypeptide is an antibody, an antigen binding fragment thereof, or an immunoadhesin,
- wherein the PEGylated binding polypeptide comprising at least one oxidized glycan is prepared by reacting a binding polypeptide comprising at least one glycan with a periodate oxidase or galactose oxidase in the presence of a salt comprising a copper metal ion to generate the PEGylated binding polypeptide comprising at least one oxidized glycan.
A method of making a PEGylated binding polypeptide, wherein the method comprises:
- (a) reacting a binding polypeptide comprising at least one glycan with a periodate oxidase or galactose oxidase in the presence of a salt comprising a copper metal ion to generate a binding polypeptide comprising at least one oxidized glycan, and
- (b) conjugating the oxidized glycan to at least one amino-oxy or hydrazine containing PEG moiety, thereby generating the PEGylated binding polypeptide, wherein the binding polypeptide is an antibody, an antigen binding fragment thereof, or an immunoadhesin.
Dependencies are inferred from the claim text.