Modified lipopolysaccharide glycoform and method of use
AssigneeThe Trustees of Princeton University
InventorsThomas J. Silhavy, Marcin Grabowicz, Daniel Kahne, Matthew Lebar, Dorothee Andres
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Abstract
The present disclosure generally relates to genetic engineering of bacteria. More particularly, the present disclosure describes genetic engineering of E. coli to create mutant O-antigen ligase, as well as novel lipopolysaccharide molecules resulting from that genetic engineering. Methods for using those novel molecules are also described.
Claims2
A vaccine adjuvant comprising at least one of a lipopolysaccharide isolated from a mutant O-antigen ligase or lipopolysaccharide derivative isolated from the mutant O-antigen ligase, the mutant O-antigen ligase comprising either an isolated protein that has the amino acid sequence SEQ ID NO:
- 1, or
- an isolated protein having at least 90% sequence identity to SEQ ID NO:
- 2 and having an amino acid substitution of phenylalanine to serine at the phenylalanine homologous to position 332 of SEQ ID NO:
- 2,
- wherein the isolated protein has at least one of 0-antigen ligase activity or the activity to ligate lipid A to disaccharide pentapeptide (DPP).
A vaccine adjuvant comprising a lipopolysaccharide derivative molecule prepared by the process comprising the steps of:
- providing an E. coli strain adapted to express an LPS derivative molecule;
- modifying the E. coli strain by creating a mutant 0-antigen ligase selected from the group consisting of:
- a mutant O-antigen ligase comprising an isolated protein that has the amino acid sequence SEQ ID NO:
- 1, and
- a mutant O-antigen ligase comprising an isolated protein having at least 90% sequence identity to SEQ ID NO:
- 2 and having an amino acid substitution of phenylalanine to serine at the phenylalanine homologous to position 332 of SEQ ID NO:
- 2, wherein the protein has at least one of 0-antigen ligase activity or the activity to ligate lipid A to disaccharide pentapeptide (DPP);
- providing a growth media for the modified E. coli strain;
- and
- allowing the modified E. coli strain to produce an LPS* derivative molecule, wherein the LPS* derivative molecule includes at least one non-native sugar not present in an LPS molecule it derives from.
Dependencies are inferred from the claim text.