Endomannosidases in the modification of glycoproteins in eukaryotes
AssigneeGlycoFi, Inc.
InventorStephen Hamilton
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Abstract
The present invention generally relates to methods of modifying the glycosylation structures of recombinant proteins expressed in fungi or other lower eukaryotes, to more closely resemble the glycosylation of proteins from higher mammals, in particular humans. The present invention also relates to novel enzymes and, nucleic acids encoding them and, hosts engineered to express the enzymes, methods for producing modified glycoproteins in hosts and modified glycoproteins so produced.
Claims25
An isolated polynucleotide comprising or consisting of a nucleic acid sequence selected from the group consisting of:
- (a) SEQ ID NO:
- 1 or 3;
- (b) a nucleic acid sequence that is a degenerate variant of SEQ ID NO:
- 1 or 3;
- (c) a nucleic acid sequence at least 78% identical to SEQ ID NO:
- 1 or 3;
- (d) a nucleic acid sequence that encodes a polypeptide having the amino acid sequence of SEQ ID NO:2 or 4;
- (e) a nucleic acid sequence that encodes a polypeptide at least 77% identical to SEQ ID NO:2 or 4;
- (f) a nucleic acid sequence that hybridizes under stringent conditions to SEQ ID NO:1 or 3;
- and
- (g) a nucleic acid sequence comprising a fragment of any one of (a)-(f) that is at least 60 contiguous nucleotides in length.
An isolated polynucleotide comprising or consisting of a nucleic acid sequence selected from the group consisting of:
- (a) SEQ ID NO:
- 1 or 3;
- (b) a nucleic acid sequence that is a degenerate variant of SEQ ID NO:
- 1 or 3;
- (c) a nucleic acid sequence at least 87% identical to SEQ ID NO:
- 1 or 3;
- (d) a nucleic acid sequence that encodes a polypeptide having the amino acid sequence of SEQ ID NO:
- 2 or 4;
- (e) a nucleic acid sequence that encodes a polypeptide at least 83% identical to SEQ ID NO:
- 2 or 4;
- (f) a nucleic acid sequence that hybridizes under stringent conditions to SEQ ID NO:
- 1 or 3;
- and
- (g) a nucleic acid sequence comprising a fragment of any one of (a)-(f) that is at least 60 contiguous nucleotides in length.
A method for modifying glycosylation structures in a lower eukaryote comprising:
- expressing an endomannosidase activity wherein the endomannosidase activity removes a composition comprising at least a glucose residue and one mannose residue on an oligosaccharide.
A method for modifing glucosylated glycoproteins comprising introducing an endomannosidase activity in a lower eukaryotic host cell wherein upon expression of the endomannosidase activity modifies a glucosylated glycoprotein that has bypassed the ER.
Dependencies are inferred from the claim text.