Alpha 1-3 n-galactosyltransferase with altered donor specificities
AssigneeHealth and Human Services
InventorsPradman K. Qasba, Boopathy Ramakrishnan, Elizabeth Boeggeman, Marta Pasek
Life of this patent
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Abstract
The invention generally features compositions and methods based on the structure-based design of alpha 1-3 N-Acetylgalactosaminyltransferase (alpha 3 GaINAc-T) enzymes from alpha 1-3 galactosyltransferase (a3Gal-T) that can transfer 2′-modified galactose from the corresponding UDP-derivatives due to substitutions that broaden the alpha 3Gal-T donor specificity and make the enzyme a3 GaINAc-T.
Claims33
A polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha 3GalNaCT) that retains the ability to transfer a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor.
A polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha3GalNAc-T) that transfers a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor, wherein the polypeptide fragment comprises and one of SEQ ID NO:
- 2, SEQ ID NO:
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- 10, SEQ ID NO;
- 12, SEQ ID NO:
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- 16, SEQ ID NO:
- 18, or SEQ ID NO:
- 20 or
- a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha3GalNac-T) that retains that ability to transfer a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor and catalyzes the formation of an oligosaccharide.
13-21. (canceled)
An isolated nucleic acid molecule comprising a nucleotide sequence which is at least 60% homologous to the nucleotide sequence of SEQ ID NO:
- 1, SEQ ID NO:
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- 17, or SEQ ID NO:
- 19 or a complement thereof, or
- an isolated nucleic acid molecule which encodes a polypeptide comprising an amino acid sequence at least about 50% homologous to the amino acid sequence of SEQ ID NO:
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- 18, or SEQ ID NO:
- 20
24-27. (canceled)
A method of making an oligosaccharide comprising incubating a reaction mixture comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha 3GalNaCT) that retains the ability to transfer a sugar with a chemically reactive functional group with a sugar donor and a sugar acceptor, or
- a method of making an oligosaccharide comprising incubating a reaction mixture comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha 3GalNaCT) that retains the ability to transfer a sugar with a chemically reactive functional group with a sugar donor and a sugar acceptor, wherein the polypeptide fragment comprises any one of SEQ ID NO:
- 2, SEQ ID NO:
- 4, SEQ ID NO:
- 6, SEQ ID NO:
- 8, SEQ ID NO:
- 10, SEQ ID NO;
- 12, SEQ ID NO:
- 14, SEQ ID NO:
- 16, SEQ ID NO:
- 18, or SEQ ID NO:
- 20.
29-48. (canceled)
An oligosaccharide synthesized by the method comprising incubating a reaction mixture comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha 3GalNAc T) that retains the ability to transfer a sugar with a chemically reactive functional group with a sugar donor and a sugar acceptor, or
- an oligosaccharide synthesized by the method comprising incubating a reaction mixture comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha 3GalNAc T) that retains the ability to transfer a sugar with a chemically reactive functional group with a sugar donor and a sugar acceptor, wherein the polypeptide fragment comprises any one of SEQ ID NO:
- 2, SEQ ID NO:
- 4, SEQ ID NO:
- 6, SEQ ID NO:
- 8, SEQ ID NO:
- 10, SEQ ID NO;
- 12, SEQ ID NO:
- 14, SEQ ID NO:
- 16, SEQ ID NO:
- 18, or SEQ ID NO:
- 20.
50-69. (canceled)
A composition comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminytransferase (alpha 3GalNaCT) that retains the ability to transfer a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor, or
- a composition comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha3GalNAc-T) that transfers a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor, wherein the polypeptide fragment comprises and one of SEQ ID NO:
- 2, SEQ ID NO:
- 4, SEQ ID NO:
- 6, SEQ ID NO:
- 8, SEQ ID NO:
- 10, SEQ ID NO;
- 12, SEQ ID NO:
- 14, SEQ ID NO:
- 16, SEQ ID NO:
- 18, or SEQ ID NO:
- 20,
- or
- a composition comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha3GalNAc-T) that retains that ability to transfer a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor and catalyzes the formation of an oligosaccharide.
71-91. (canceled)
An immunological composition comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha 3GalNAc-T) that retains the ability to transfer a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor, or
- an immunological composition comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha3GalNAc-T) that transfers a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor, wherein the polypeptide fragment comprises and one of SEQ ID NO:
- 2, SEQ ID NO:
- 4, SEQ ID NO:
- 6, SEQ ID NO:
- 8, SEQ ID NO:
- 10, SEQ ID NO;
- 12, SEQ ID NO:
- 14, SEQ ID NO:
- 16, SEQ ID NO:
- 18, or SEQ ID NO:
- 20, and wherein one or more antibodies are conjugated to the chemically reactive functional group, or
- an immunological composition comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha3GalNAc-T) that retains that ability to transfer a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor and catalyzes the formation of an oligosaccharide, and wherein one or more antibodies are conjugated to the chemically reactive functional group.
93-113. (canceled)
A method of coupling an agent to a carrier protein comprising:
- incubating a reaction mixture comprising a polypeptide fragment of an alpha 1,3 N-acetylgalactosaminyltransferase (alpha 3GalNAc T) that retains the ability to transfer a sugar with a chemically reactive functional group from a sugar donor to a sugar acceptor, wherein the sugar donor is coupled to an agent and the sugar acceptor is a carrier protein.
115-137. (canceled)
A method for the diagnosis or treatment of a subject suffering from a disease or disorder comprising:
- administering to the subject an effective amount of an isolated glycoprotein synthesized by the method according to claim 60, wherein one or more agents are linked to the sugar donor,
- thereby diagnosing or treating a subject suffering from a disease or disorder.
139-140. (canceled)
A method for imaging a target cell or tissue comprising:
- administering to a subject an oligosaccharide synthesized by the method according to claim 48, and wherein one or more imaging agents are linked to the sugar donor, thereby imaging a target cell or tissue.
A method for synthesizing a detectable Gal beta 1-4GlcNAc epitope comprising:
- synthesizing an oligosaccharide according to the method of claim 50, wherein the sugar donor comprises a 2′ modified Gal residue and wherein or more detection agents are linked to the 2′ modified Gal residue thereby synthesizing a detectable Gal beta 1-4GlcNAc epitope.
143-145. (canceled)
147-150. (canceled)
Dependencies are inferred from the claim text.