Modified host cells and hybrid oligosaccharides for use in bioconjugate production
AssigneeGlycovaxyn AG
InventorAmirreza FARIDMOAYER
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Abstract
Provided herein are host cells capable of producing hybrid oligosaccharides and polysaccharides, wherein said hybrid oligosaccharides and polysaccharides do not comprise a hexose at the reducing end of their first repeat unit. Also provided herein are hybrid oligosaccharides or polysaccharides and bioconjugates which can be produced by the host cells described herein, wherein said bioconjugates comprise a carrier protein linked to a hybrid oligosaccharide or polysaccharide that does not comprise a hexose at the reducing end of its first repeat unit.
Claims29
1-96. (canceled)
A prokaryotic host cell comprising:
- a) nucleic acids that encode (i) glycosyltransferases that produce an oligosaccharide or polysaccharide repeat unit, wherein said repeat unit does not comprise a hexose at the reducing end, and wherein said oligosaccharide or polysaccharide repeat unit is derived from a donor oligosaccharide or polysaccharide repeat unit that comprises a hexose at the reducing end, and (ii) glycosyltransferases sufficient for synthesis of the donor oligosaccharide or polysaccharide repeat unit;
- and
- b) a nucleic acid that encodes a carrier protein comprising an N-glycosylation consensus sequence;
- and
- c) a nucleic acid that encodes an oligosaccharyl transferase;
- and
- d) a nucleic acid that encodes a wzy polymerase, wherein the polymerase catalyzes the production of a hybrid oligosaccharide or polysaccharide, wherein the hybrid oligosaccharide or polysaccharide (i) comprises a hexose monosaccharide derivative at the reducing end of the first repeat unit and (ii) comprises hexose monosaccharides at the reducing end of all other repeat units.
A hybrid oligosaccharide or polysaccharide having a structure
- (B)n-A→
- a) wherein A is an oligosaccharide repeat unit (i) of a capsular saccharide of a Gram positive bacterial capsular saccharide selected from a Group A streptococcus capsular saccharide, a Group B streptococcus capsular saccharide, a Streptococcus pneumoniae capsular saccharide, Enterococcal capsular saccharide or a Staphylococcus aureus capsular saccharide, (ii) that contains at least 2, 3, 4, 5, 6, 7 or 8 monosaccharides, (iii) with a hexose monosaccharide derivative at the reducing end (indicated by arrow);
- b) wherein B is an oligosaccharide repeat unit (i) containing at least 2, 3, 4, 5, 6, 7 or 8 monosaccharides and (ii) with a hexose monosaccharide at the reducing end of the repeat optionally wherein the hexose monosaccharide comprises glucose, galactose, rhamnose, arabinotol, fucose, or mannose;
- c) wherein A and B are different oligosaccharide repeat units that optionally differ only by containing a different monosaccharide at the reducing end of the repeat;
- and
- wherein n is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
Dependencies are inferred from the claim text.