Abstract: The present invention discloses a conjugate comprising an antigen (for example a saccharide antigen) covalently linked to a Pseudomonas aeruginosa PcrV carrier protein comprising an amino acid sequence which is at least 80% identical to the sequence of SEQ ID NO:1-4, wherein the antigen is linked (either directly or through a linker) to an amino acid residue of the P. aeruginosa PcrV carrier protein. The invention also discloses Pseudomonas aeruginosa PcrV proteins that contain glycosylation site consensus sequences.

Abstract: The present invention discloses a process for engineering a host cell comprising the steps of; a) integrating a first polynucleotide cassette including a first selection marker flanked by a first pair of recombination sites; b) removing the first selection marker by the action of a recombinase which recognises the first pair of recombination sites; c) integrating a second polynucleotide cassette including a second selection marker flanked by a second pair of recombination sites; and d) removing the second selection marker by the action of a recombinase which recognises the second pair of recombination sites. Also disclosed is a host cell genome polynucleotide comprising a first recombinantly engineered region and a second recombinantly engineered region, wherein a first single recombination site is adjacent to the first recombinantly engineered region, and a second single recombination site is adjacent to the second recombinantly engineered region.

Abstract: The disclosure provides synthetic (e.g. recombinant) pneumococcal saccharide comprising one or more repeat unit(s) ?4)-?-D-Glcp-(1?3)-[[?-L-Rhap-(1?2)]-[Gro-(2?P?3)]-?-D-Galp-(1?4)]-?-L-Rhap-(1?. Also provided are conjugates comprising a ?4)-?-D-Glcp-(1?3)-[[?-L-Rhap-(1?2)]-[Gro-(2?P?3)]-?-D-Galp-(1?4)]-?-L-Rhap-(1?, immunogenic compositions, vaccines and their use in preventing or treating infection by Streptococcus pneumoniae.

Abstract: A bioconjugate of an E. coli glucosylated O4 antigen polysaccharide covalently linked to a carrier protein and compositions thereof are provided. Also provided are recombinant host cells for producing the bioconjugate, and methods of producing the bioconjugate using the recombinant host cells. The recombinant host cells contain a nucleic acid encoding a glucosyl transferase capable of modifying the E. coli O4 antigen with glucose branching to produce the glucosylated O4 antigen polysaccharide. Bioconjugates of an E. coli glucosylated O4 antigen polysaccharide described herein can be used alone or in combination with one or more additional E. coli O-antigen polysaccharides to induce antibodies against an E. coli glucosylated antigen, and to vaccinate a subject against extra-intestinal pathogenic E. coli (ExPEC).

Abstract: The present invention relates to the field of immunogenic compositions and vaccines, their manufacture, host cells which can be used in their manufacture and the use of such immunogenic compositions and vaccines in medicine. More particularly, it relates to Klebsiella pneumoniae O-antigens, conjugates comprising a K. pneumoniae O-antigen, host cells suitable for their production and immunogenic compositions or vaccines containing at least one Klebsiella pneumoniae O-antigen.

Abstract: Methods of producing bioconjugates of O-antigen polysaccharides covalently linked to a carrier protein using recombinant host cells are provided. The recombinant host cells used in the methods described herein encode a particular oligosaccharyl transferase enzyme depending on the O-antigen polysaccharide bioconjugate to be produced. The oligosaccharyl transferase enzymes can be PglB oligosaccharyl transferase or variants thereof. Also provided are compositions containing the bioconjugates, and methods of using the bioconjugates and compositions described herein to vaccinate a subject against extra-intestinal pathogenic E. coli. (ExPEC).

Abstract: The disclosure provides synthetic (e.g. recombinant) pneumococcal saccharide comprising one or more repeat unit(s) ?4)-?-D-Glcp-(1?3)-[[?-L-Rhap-(1?2)]-[Gro-(2?P?3)]-?-D-Galp-(1?4)]-?-L-Rhap-(1?. Also provided are conjugates comprising a ?4)-?-D-Glcp-(1?3)-[[?-L-Rhap-(1?2)]-[Gro-(2?P?3)]-?-D-Galp-(1?4)]-?-L-Rhap-(1?, immunogenic compositions, vaccines and their use in preventing or treating infection by Streptococcus pneumoniae.

Abstract: A bioconjugate of an E. coli glucosylated O4 antigen polysaccharide covalently linked to a carrier protein and compositions thereof are provided. Also provided are recombinant host cells for producing the bioconjugate, and methods of producing the bioconjugate using the recombinant host cells. The recombinant host cells contain a nucleic acid encoding a glucosyl transferase capable of modifying the E. coli O4 antigen with glucose branching to produce the glucosylated O4 antigen polysaccharide. Bioconjugates of an E. coli glucosylated O4 antigen polysaccharide described herein can be used alone or in combination with one or more additional E. coli O-antigen polysaccharides to induce antibodies against an E. coli glucosylated antigen, and to vaccinate a subject against extra-intestinal pathogenic E. coli (ExPEC).

Abstract: Compositions and methods are described for inducing an immune response against extra-intestinal pathogenic Escherichia coli (ExPEC) to thereby provide immune protection against diseases associated with ExPEC. In particular, compositions and methods are described for using conjugates of E. coli polysaccharide antigens O25B, O1A, O2, and O6A covalently bound to a detoxified exotoxin A of Pseudomonas aeruginosa (EPA) carrier protein as vaccines for the prevention of invasive ExPEC disease caused by ExPEC serotypes O1A, O2, O6A and O25B.

Abstract: The disclosure provides synthetic (e.g. recombinant) pneumococcal saccharides comprising one or more repeat unit(s) ?4)-?-D-Glcp-(1?4)-[Gro-(2?P?3)]-?-D-Galp-(1?4)-?-L-Rhap-(1?. Also provided are conjugates comprising a ?4)-?-D-Glcp-(1?4)-[Gro-(2?P?3)]-?-D-Galp-(1?4)-?-L-Rhap-(1?, immunogenic compositions, vaccines and their use in preventing or treating infection by Streptococcus pneumoniae.

Abstract: The present disclosure provides mutated PglB oligosaccharyltransferase enzymes that have the ability to efficiently catalyze the transfer of a saccharide from a lipid carrier to an asparagine reissue in a glycosylation motif on a protein. Also provided are polynucleotides encoding the mutated PglB oligosaccharyltransferase enzymes, host cells capable of expressing the engineered PglB oligosaccharyltransferase enzymes, and methods of using the engineered PglB oligosaccharyltransferase enzymes to make N-glycosylated proteins. Also provided are N-glycosylated proteins that are made using the engineered PglB oligosaccharyltransferase enzymes.

Abstract: Methods of producing bioconjugates of O-antigen polysaccharides covalently linked to a carrier protein using recombinant host cells are provided. The recombinant host cells used in the methods described herein encode a particular oligosaccharyl transferase enzyme depending on the O-antigen polysaccharide bioconjugate to be produced. The oligosaccharyl transferase enzymes can be PglB oligosaccharyl transferase or variants thereof. Also provided are compositions containing the bioconjugates, and methods of using the bioconjugates and compositions described herein to vaccinate a subject against extra-intestinal pathogenic E. coli. (ExPEC).

Abstract: Compositions and methods are described for inducing an immune response against extra-intestinal pathogenic Escherichia coli (ExPEC) to thereby provide immune protection against diseases associated with ExPEC. In particular, compositions and methods are described for using conjugates of E. coli polysaccharide antigens O25B, O1A, O2, and O6A covalently bound to a detoxified exotoxin A of Pseudomonas aeruginosa (EPA) carrier protein as vaccines for the prevention of invasive ExPEC disease caused by ExPEC serotypes O1A, O2, O6A and O25B.

Abstract: A bioconjugate of an E. coli glucosylated O4 antigen polysaccharide covalently linked to a carrier protein and compositions thereof are provided. Also provided are recombinant host cells for producing the bioconjugate, and methods of producing the bioconjugate using the recombinant host cells. The recombinant host cells contain a nucleic acid encoding a glucosyl transferase capable of modifying the E. coli O4 antigen with glucose branching to produce the glucosylated O4 antigen polysaccharide. Bioconjugates of an E. coli glucosylated O4 antigen polysaccharide described herein can be used alone or in combination with one or more additional E. coli O-antigen polysaccharides to induce antibodies against an E. coli glucosylated antigen, and to vaccinate a subject against extra-intestinal pathogenic E. coli (ExPEC).

Abstract: The present invention discloses a conjugate comprising an antigen (for example a saccharide antigen) covalently linked to a Pseudomonas aeruginosa PcrV carrier protein comprising an amino acid sequence which is at least 80% identical to the sequence of SEQ ID NO:1-4, wherein the antigen is linked (either directly or through a linker) to an amino acid residue of the P. aeruginosa PcrV carrier protein. The invention also discloses Pseudomonas aeruginosa PcrV proteins that contain glycosylation site consensus sequences.

Abstract: The disclosure provides synthetic (e.g. recombinant) pneumococcal saccharides comprising one or more repeat unit(s) ?4)-?-D-Glcp-(1?4)-[Gro-(2?P?3)]-?-D-Galp-(1?4)-?-L-Rhap-(1?. Also provided are conjugates comprising a ?4)-?-D-Glcp-(1?4)-[Gro-(2?P?3)]-?-D-Galp-(1?4)-?-L-Rhap-(1?, immunogenic compositions, vaccines and their use in preventing or treating infection by Streptococcus pneumoniae.

Abstract: The present invention discloses a process for engineering a host cell comprising the steps of; a) integrating a first polynucleotide cassette including a first selection marker flanked by a first pair of recombination sites; b) removing the first selection marker by the action of a recombinase which recognises the first pair of recombination sites; c) integrating a second polynucleotide cassette including a second selection marker flanked by a second pair of recombination sites; and d) removing the second selection marker by the action of a recombinase which recognises the second pair of recombination sites. Also disclosed is a host cell genome polynucleotide comprising a first recombinantly engineered region and a second recombinantly engineered region, wherein a first single recombination site is adjacent to the first recombinantly engineered region, and a second single recombination site is adjacent to the second recombinantly engineered region.

Abstract: Carrier proteins modified to incorporate one or more pilin glycotags and applications thereof for O-linked glycosylation are provided. In particular, a modified carrier protein comprising a carrier protein that comprises at least one GlycoTag, wherein the at least one GlycoTag is a Neisseria gonorrhoeae PglL GlycoTag (NgGlycoTag), Neisseria lactamica PglL GlycoTag (NlGlycoTag), or Neisseria shayeganii GlycoTag (NsGlycoTag), or combinations thereof is provided, together with nucleic acids and vectors encoding the modified carrier protein, host cells comprising these modofoed carrier proteins or nucleic acids encoding them, bioconjugates, methods of making bioconjugates and uses of the bioconjugates.

Abstract: A process for the production of a glycoconjugate by N-glycosylation of a protein or peptide comprising the sequence D/E-X-N-X-S/T (SEQ ID NO:1), wherein each X is the same or different and is any natural amino acid other than proline, wherein the process comprises reacting the protein or peptide with a polyisoprenyl pyrophosphate of formula (I), or a salt thereof, in the presence of PglB: to produce the glycoconjugate comprising the protein or peptide having a saccharide [S1] linked to the asparagine in the sequence D/E-X-N-X-S/T (SEQ ID NO:1). Polyisoprenylpyrophosphates used as substrates in the biocatalytic process are also provided, as well as certain glycoconjugates.

Abstract: Described herein are oligosaccharyl transferases for use in N-glycosylating proteins of interest in vitro and in host cells. Methods for using such oligosaccharyl transferases, nucleic acids encoding such oligosaccharyl transferases, and host cells comprising such oligosaccharyl transferases are also provided herein. Glycoconjugates generated by using such oligosaccharyl transferases are also provided herein.