	US 12,233,121 B2
	*Methods of producing bioconjugates of E. coli* O-antigen polysaccharides, compositions thereof, and methods of use thereof**
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 (US); Stefan Jochen Kemmler, Zurich (CH); Michael Thomas Kowarik, Zurich (CH); Manuela Mally, Watt (CH); Veronica Gambillara Fonck, Meilen (CH); Martin Edward Braun, Cham (CH); and Maria Paula Carranza Sandmeier, Rudolfstetten (CH)
Assigned to Janssen Pharmaceuticals, Inc., Titusville, NJ (US); and GlaxoSmith Kline Biologicals S.A., Rixensart (BE)
Filed by Janssen Pharmaceuticals, Inc., Titusville, NJ (US); and GlaxoSmithKline Biologicals S.A., Rixensart (BE)
Filed on Oct. 6, 2022, as Appl. No. 17/938,454.
Application 17/938,454 is a division of application No. 16/822,403, filed on Mar. 18, 2020, granted, now 11,491,220.
Claims priority of provisional application 62/819,762, filed on Mar. 18, 2019.
Prior Publication US 2023/0190926 A1, Jun. 22, 2023
Int. Cl. A61K 39/108 (2006.01); A61K 39/00 (2006.01); A61K 39/02 (2006.01); A61K 39/085 (2006.01); A61K 39/09 (2006.01); A61K 39/095 (2006.01); A61K 39/102 (2006.01); A61K 39/104 (2006.01); A61K 47/64 (2017.01)

CPC A61K 39/104 (2013.01) [A61K 39/0258 (2013.01); A61K 39/085 (2013.01); A61K 39/092 (2013.01); A61K 39/095 (2013.01); A61K 39/102 (2013.01); A61K 39/107 (2013.01); A61K 47/646 (2017.08); A61K 2039/6037 (2013.01); A61K 2039/62 (2013.01)]

8 Claims

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.