US 12,290,563 B2
Production of E. coli 018 bioconjugates
Jeroen Geurtsen, Vleuten (NL); Marleen Eveline Weerdenburg, Uithoorn (NL); and Pieter Jan Burghout, Pijnacker (NL)
Assigned to Janssen Pharmaceuticals, Inc., Titusville, NJ (US)
Filed by Janssen Pharmaceuticals, Inc., Titusville, NJ (US)
Filed on Mar. 31, 2022, as Appl. No. 17/710,116.
Claims priority of application No. 21166781 (EP), filed on Apr. 1, 2021.
Prior Publication US 2022/0323576 A1, Oct. 13, 2022
Int. Cl. A61K 39/385 (2006.01); A61K 39/02 (2006.01); A61K 39/108 (2006.01); A61K 47/64 (2017.01); A61P 31/04 (2006.01); A61P 37/04 (2006.01); C12N 9/10 (2006.01); A61K 39/00 (2006.01)
CPC A61K 39/385 (2013.01) [A61K 39/0258 (2013.01); A61K 47/646 (2017.08); A61P 31/04 (2018.01); A61P 37/04 (2018.01); C12N 9/1081 (2013.01); A61K 2039/55583 (2013.01); A61K 2039/6087 (2013.01); A61K 2039/70 (2013.01)] 19 Claims
 
1. A gram-negative bacterial host cell comprising:
(a) a carrier protein comprising at least one glycosylation consensus sequence;
(b) an E. coli O18 rfb locus; and,
(c) an oligosaccharyltransferase;
wherein the cell comprises a polypeptide having Wzy O-antigen polymerase activity, which polypeptide comprises an amino acid sequence with at least 95% identity with SEQ ID NO: 1, wherein the amino acid sequence comprises:
i) isoleucine at a position that corresponds to position 199 in SEQ ID NO: 1, lysine at a position that corresponds to position 377 in SEQ ID NO: 1, and alanine at a position that corresponds to position 395 in SEQ ID NO: 1;
ii) threonine at a position that corresponds to position 199 in SEQ ID NO: 1, lysine at a position that corresponds to position 377 in SEQ ID NO: 1, and valine at a position that corresponds to position 395 in SEQ ID NO: 1;
iii) threonine at a position that corresponds to position 199 in SEQ ID NO: 1, lysine at a position that corresponds to position 377 in SEQ ID NO: 1, and alanine at a position that corresponds to position 395 in SEQ ID NO: 1;
iv) isoleucine at a position that corresponds to position 199 in SEQ ID NO: 1, lysine at a position that corresponds to position 377 in SEQ ID NO: 1, and valine at a position that corresponds to position 395 in SEQ ID NO: 1; or
v) isoleucine at a position that corresponds to position 199 in SEQ ID NO: 1, methionine at a position that corresponds to position 377 in SEQ ID NO: 1, and alanine at a position that corresponds to position 395 in SEQ ID NO: 1.