Granted Patent B2

US 12,233,121 · App. 17/938,454 · Granted Feb 25, 2025

Inventors: Jeroen Geurtsen (Vleuten, NL); Pieter Jan Burghout (Pijnacker, NL); Eveline Marleen Weerdenburg (Uithoorn, NL); Jan Theunis Poolman (Vogelenzang, NL); Kellen Cristhina Fae (Mainz, DE); Patricia Ibarra Yon (Solothum, CH); Darren Robert Abbanat (Cornwall, NY); Stefan Jochen Kemmler (Zurich, CH); Michael Thomas Kowarik (Zurich, CH); Manuela Mally (Watt, CH); Veronica Gambillara Fonck (Meilen, CH); Martin Edward Braun (Cham, CH); Maria Paula Carranza Sandmeier (Rudolfstetten, CH)

Assignees: Janssen Pharmaceuticals, Inc.; GlaxoSmith Kline Biologicals S.A.

A61K39/104A61K39/0258A61K39/085A61K39/092A61K39/095A61K39/102A61K39/107A61K47/646A61K2039/6037A61K2039/62

Abstract

Claims (14)

1. A recombinant host cell for preparing a bioconjugate of an E. coli Ox-antigen polysaccharide covalently linked to a carrier protein, the recombinant host cell comprising:

(a) a nucleotide sequence of an rfb gene cluster for the Ox-antigen polysaccharide;

(b) a nucleotide sequence encoding the carrier protein comprising at least one glycosylation site comprising a glycosylation consensus sequence having SEQ ID NO: 1; and

(c) a nucleotide sequence encoding an oligosaccharyl transferase PglBy, wherein the PglBy comprises the amino acid substitution N311V relative to a wild-type PglB having the amino acid sequence of SEQ ID NO: 6,

wherein the E. coli Ox-antigen polysaccharide is the O1A antigen polysaccharide comprising the structure:

wherein n is independently an integer of 3 to 50.

2. The recombinant host cell of claim 1 , wherein

the PglB y comprises the amino acid mutations of N311V, K482R, D483H, and A669V relative to the wild-type PglB.

3. The recombinant host cell of claim 1 , wherein the host cell is a prokaryotic host cell.

4. The recombinant host cell of claim 3 , wherein the host cell is E. coli.

5. The recombinant host cell of claim 1 , wherein the carrier protein is selected from the group consisting of detoxified Exotoxin A of P. aeruginosa (EPA), E. coli flagellin (FliC), CRM197, maltose binding protein (MBP), Diphtheria toxoid, Tetanus toxoid, detoxified hemolysin A of S. aureus , clumping factor A, clumping factor B, E. coli heat labile enterotoxin, detoxified E. coli heat labile enterotoxin, cholera toxin B subunit (CTB), cholera toxin, detoxified cholera toxin, E. coli Sat protein, the passenger domain of E. coli Sat protein, Streptococcus pneumoniae Pneumolysin, Keyhole limpet hemocyanin (KLH), P. aeruginosa PcrV, outer membrane protein of Neisseria meningitidis (OMPC), and protein D from non-typeable Haemophilus influenzae.

6. The recombinant host cell of claim 1 , wherein the carrier protein comprises SEQ ID NO: 3.

7. The recombinant host cell of claim 1 , wherein the PglBy comprises the amino acid sequence of SEQ ID NO: 6 with the N311V substitution, the host cell is E. coli and the carrier protein comprises the amino acid sequence of SEQ ID NO: 3.

8. The recombinant host cell of claim 1 , wherein the PglBy comprises the amino acid sequence of SEQ ID NO: 6 with the N311V, K482R, D483H and A669V substitutions, the host cell is E. coli and the carrier protein comprises the amino acid sequence of SEQ ID NO: 3.

Assignments (8)

Continuity (3)

References Cited (226)