# US 2019/0169216 A1 · Purification of staphylococcus aureus type 5 and type 8 capsular saccharides > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0169216 A1 - Document ID: US20190169216A1 - Status: Published application - Kind: A1 - Application number: 16/274,822 - Filed: 2019-02-13 - Published: 2019-06-06 - Earliest claimed: 2009-10-30 - Assignees: Glaxosmithkline Biologicals SA - Applicants: Glaxosmithkline Biologicals, S.A. - Inventors: Paolo COSTANTINO; Maria Rosaria ROMANO; Francesco BERTI - Web page: https://patentgrep.com/patent/US20190169216A1 ## CPC - C07H 1/08: from natural products - A61K 39/085: Staphylococcus - A61P 31/04: Antibacterial agents - A61P 37/04: Immunostimulants - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - C12P 19/04: Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds - A61K 2039/55505: {Inorganic adjuvants} - A61K 2039/6037: {Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT]} - C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)} ## Abstract The invention provides a method for releasing capsular polysaccharide from S. aureus type 5 or type 8 cells, comprising the step of treating the cells with acid. The invention further provides a process for purifying capsular polysaccharide from S. aureus type 5 or type 8 cells comprising this method. Other processing steps may be included in the process, such as enzymatic treatment, e.g. to remove nucleic acid, protein and/or peptidoglycan contaminants; diafiltration, e.g. to remove low molecular weight contaminants; anion exchange chromatography, e.g. to remove residual protein; and concentration. ## Claims ### Claim at position 1 1: A method for releasing capsular polysaccharide from S. aureus type 5 or 8 cells, comprising the step of treating the cells with acid. ### Claim at position 2 (depends on 1) 2: The method of claim 1, wherein the cells are in the form of a wet cell paste or are suspended in an aqueous medium. ### Claim at position 3 (depends on 1) 3: The method of claim 1, wherein the acid treatment is carried out using acetic acid. ### Claim at position 4 (depends on 1) 4: The method of claim 1, wherein the acid treatment results in the capsular polysaccharide having a degree of O-acetylation between 60-100%. ### Claim at position 5 (depends on 1) 5: The method of claim 1, wherein the method further comprises a step of neutralisation. ### Claim at position 6 (depends on 1) 6: The method of claim 1, wherein the method further comprises a step of centrifugation of the cells and collection of the polysaccharide-containing supernatant. ### Claim at position 7 (depends on 1) 7: A process for purifying capsular polysaccharide from S. aureus type 8 cells comprising the method of claim 1. ### Claim at position 8 (depends on 7) 8: The process of claim 7, wherein the process further comprises a step of treatment of the capsular polysaccharide with DNase and/or RNase. ### Claim at position 9 (depends on 7) 9: The process of claim 7, wherein the process further comprises a step of treatment of the capsular polysaccharide with mutanolysin. ### Claim at position 10 (depends on 7) 10: The process of claim 7, wherein the process further comprises a step of diafiltration. ### Claim at position 11 (depends on 10) 11: The process of claim 10, wherein the diafiltration is tangential flow filtration. ### Claim at position 12 (depends on 7) 12: The process of claim 7, wherein the process further comprises a step of anion exchange chromatography. ### Claim at position 13 (depends on 7) 13: The process of claim 7, wherein the process further comprises a step of gel filtration. ### Claim at position 14 (depends on 7) 14: The process of claim 7, wherein the process further comprises a step of concentration of the polysaccharide. ### Claim at position 15 (depends on 7) 15: The process of claims 7, wherein the molecular mass of the purified polysaccharide is between 2-3500 kDa. ### Claim at position 16 (depends on 7) 16: The process of claim 7, wherein the process further comprises a step of depolymerisation of the purified polysaccharide to form an oligosaccharide. ### Claim at position 17 (depends on 7) 17: The process of claim 7, wherein the process further comprises a step of sterile filtration. ### Claim at position 18 (depends on 7) 18: The process of claim 7, wherein the process provides a composition comprising the polysaccharide and a level of peptidoglycan contamination that is less than 5% by weight peptidoglycan relative to the total weight of the polysaccharide. ### Claim at position 19 (depends on 18) 19: The process of claim 18, wherein the level of peptidoglycan contamination is about 2%. ### Claim at position 20 (depends on 7) 20: The process of claim 7, wherein the process provides a composition comprising the polysaccharide and a level of protein contamination that is less than 5% by weight protein relative to the total weight of the polysaccharide. ### Claim at position 21 (depends on 7) 21: The process of claim 7, wherein the process provides a composition comprising the polysaccharide and a level of nucleic acid contamination that is less than 1% by weight nucleic acid relative to the total weight of the polysaccharide. ### Claim at position 22 (depends on 7) 22: The process of claim 7, wherein the process further comprises a step of conjugation to a carrier molecule. ### Claim at position 23 (depends on 7) 23: A composition comprising an S. aureus type 5 or type 8 capsular polysaccharide obtainable by the process of claims 7 or a conjugate obtainable by the process of claim ### Claim at position 24 (depends on 23) 24: The composition of claim 23 further comprising one or more S. aureus protein antigen(s) selected from the group consisting of a clfA antigen; a clfB antigen; a sdrE2 antigen; a sdrC antigen; a sasF antigen; a emp antigen; a sdrD antigen; a spa antigen; a esaC antigen; a esxA antigen; a esxB antigen; a sta006 antigen; a isdC antigen; a Hla antigen; a sta011 antigen; a isdA antigen; a isdB antigen; and a sta073 antigen. ### Claim at position 25 (depends on 24) 25: The composition of claim 24, wherein the one or more S. aureus protein antigen(s) are selected from the group consisting of a esxA antigen; a esxB antigen; a sta006 antigen; a Hla antigen; a sta011 antigen; and a sta073 antigen. ### Claim at position 26 (depends on 25) 26: The composition of claim 25, wherein the composition comprises S. aureus protein antigens according to one of combinations (1) to (10) below: (1) a esxA antigen, a esxB antigen, a sta006 antigen and a Hla antigen; (2) a esxA antigen, a esxB antigen, a sta006 antigen and a sta011 antigen; (3) a esxA antigen, a esxB antigen and a sta011 antigen; (4) a esxA antigen, a esxB antigen, a Hla antigen, a sta006 antigen and a sta011 antigen; (5) a esxA antigen, a esxB antigen and a Hla antigen; (6) a Hla antigen, a sta006 antigen and a sta011 antigen; (7) a esxA antigen and a esxB antigen; (8) a esxA antigen, a esxB antigen and a sta006 antigen; (9) a esxA antigen, a esxB antigen, a sta011 antigen and a sta073 antigen; and a sta006 antigen and a sta011 antigen.