System and method for the production of recombinant glycosylated proteins in a prokaryotic host
AssigneeETH Zurich
InventorsMarkus Aebi, Michael Wacker
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Abstract
A system and a method for the production of recombinant N-glycosylated target proteins. The system comprises a prokaryotic organism (e.g. Escherichia coli) into which is introduced a genetic information encoding for a metabolic apparatus capable of carrying out the requested N-glycosylation of the target protein. Said prokaryotic organism also contains the genetic information required for the expression of one or more recombinant target proteins. The metabolic apparatus preferably comprises specific glycosyltransferases for the assembly of the oligosaccharide on a lipid carrier and an OTase that covalently links this oligosaccharide to specific residues of the desired protein.
Claims7
A host prokaryotic organism comprising:
- (a) a target protein having a consensus sequence Asn-X-Ser(Thr), wherein X can be any amino acid except Pro, and
- (b) an oligosaccharyl transferase, wherein the oligosaccharyl transferase is the oligosaccharyl transferase from Campylobacter jejuni,
- wherein the target protein and the oligosaccharyl transferase are heterologous to the host organism and wherein the target protein is glycosylated at the Asn residue of the consensus sequence.
A prokaryotic expression system obtained by a process comprising introducing into a host prokaryotic organism nucleic acids encoding:
- (a) a target protein, having a consensus sequence Asn-X-Ser(Thr), wherein X can be any amino acid except Pro, and
- (b) an oligosaccharyl transferase, wherein the oligosaccharyl transferase is the oligosaccharyl transferase from Campylobacter jejuni,
- wherein the target protein and the oligosaccharyl transferase are heterologous to the host organism and wherein the target protein is glycosylated at the Asn residue of the consensus sequence.
Dependencies are inferred from the claim text.