US 11,931,405 B2
Bioconjugates of E. coli O-antigen polysaccharides, methods of production thereof, and methods of use thereof
Jeroen Geurtsen, Vleuten (NL); Jan Theunis Poolman, Vogelenzang (NL); Kellen Cristhina Fae, Mainz (DE); Pieter Jan Burghout, Pijnacker (NL); Eveline Marleen Weerdenburg, Uithoorn (NL); Patricia Ibarra Yon, Solothum (CH); Darren Robert Abbanat, Cornwal, 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.; and GlaxoSmithKline Biologicals S.A., Rixensart (BE)
Filed by Janssen Pharmaceuticals, Inc., Titusville, NJ (US); and GlaxoSmithKline Biologicals S.A., Rixensart (BE)
Filed on Jul. 28, 2022, as Appl. No. 17/815,705.
Application 17/815,705 is a division of application No. 16/822,340, filed on Mar. 18, 2020, granted, now 11,446,370.
Claims priority of provisional application 62/819,746, filed on Mar. 18, 2019.
Prior Publication US 2023/0118878 A1, Apr. 20, 2023
Int. Cl. A61K 39/108 (2006.01); A61K 39/00 (2006.01); A61P 31/04 (2006.01); A61P 37/04 (2006.01); C07K 14/21 (2006.01); C07K 19/00 (2006.01); C12N 9/10 (2006.01)
CPC A61K 39/0258 (2013.01) [A61P 31/04 (2018.01); A61P 37/04 (2018.01); C07K 14/21 (2013.01); C07K 19/00 (2013.01); C12N 9/1051 (2013.01); C12Y 204/01 (2013.01); A61K 2039/575 (2013.01); A61K 2039/6037 (2013.01)] 22 Claims
 
1. A recombinant host cell for producing a bioconjugate of an E. coli glucosylated O4 antigen polysaccharide covalently linked to a carrier protein, wherein the glucosylated O4 antigen polysaccharide comprises the structure of Formula

OG Complex Work Unit Chemistry
wherein n is an integer of 5 to 40,
the host cell comprising:
(i) a nucleotide sequence of an rfb gene cluster for the E. coli O4 antigen polysaccharide;
(ii) a nucleotide sequence encoding a glucosyl transferase, wherein the glucosyl transferase is capable of modifying the E. coli O4 antigen polysaccharide to produce the E. coli glucosylated O4 antigen polysaccharide;
(iii) a nucleotide sequence encoding a translocase that is capable of translocating bactoprenol-linked glucose;
(iv) a nucleotide sequence encoding a glycosyltransferase that is capable of glucosylating bactoprenol;
(v) a nucleotide sequence encoding the carrier protein; and
(vi) a nucleotide sequence encoding an oligosaccharyl transferase capable of covalently linking the E. coli glucosylated O4 antigen polysaccharide to the carrier protein to produce the bioconjugate.