Capsular polysaccharide solubilisation and combination vaccines
AssigneeGlaxosmithkline Biologicals SA
InventorPaolo COSTANTINO
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Abstract
Precipitated bacterial capsular polysaccharides can be efficiently re-solubilised using alcohols as solvents. The invention provides a process for purifying a bacterial capsular polysaccharide, comprising the steps of (a) precipitation of said polysaccharide, followed by (b) solubilisation of the precipitated polysaccharide using ethanol. CTAB can be used for step (a). The material obtained, preferably following hydrolysis and sizing, can be conjugated to a carrier protein and formulated as a vaccine. Also, in vaccines comprising saccharides from both serogroups A and C, the invention provides that the ratio (w/w) of Men A saccharide: MenC saccharide is >1.
Claims21
1-63. (canceled)
7:. A composition comprising a MenA capsular saccharide conjugate and a MenC capsular saccharide conjugate, and further comprising (i) an aluminum hydroxide adjuvant and (ii) a histidine buffer, and where each conjugate has a saccharide:protein ratio (w/w) of between 0.5:1 and 5:1.
64:
- An immunogenic composition comprising capsular saccharides from (a) at least serogroup A of N. meningitidis (MenA) and serogroup C of N. meningitidis (MenC), wherein the ratio of MenA:MenC is greater than 1, wherein the MenA and MenC capsular saccharides are conjugated to carrier proteins, and/or (b) serogroup Y of N. meningitidis (MenY) and one or both of serogroup C of N. meningitidis (MenC) and serogroup W135 of N. meningitidis (MenW135), wherein MenY:MenW135 is greater than 1, and/or MenY:MenC is less than 1 and wherein the MenY capsular saccharides are conjugated to carrier proteins.
65:
- The composition of claim 64, wherein the capsular saccharides are oligosaccharides.
66:
- The composition of claim 65, wherein the composition comprises MenC capsular saccharides and MenY capsular saccharides.
67:
- The composition of claim 65, wherein the composition comprises MenC capsular saccharides, MenW135 capsular saccharides, and MenY capsular saccharides.
68:
- The composition of claim 65, wherein the composition comprises MenA capsular saccharides, MenC capsular saccharides, MenW135 capsular saccharides, and MenY capsular saccharides.
69:
- The composition of claim 68, wherein the ratios (w/w) for the capsular saccharides from MenA:MenC:MenW135:MenY are:
- 1:1:1:2;
- 2:1:1:1;
- 4:2:1:1;
- 8:4:2:1;
- 4:2:1:2;
- 8:4:1:2;
- 4:2:2:1;
- 2:2:1:1;
- 4:4:2:1;
- 2:2:1:2;
- 4:4:1:2;
- or 2:2:2:1.
70:
- The composition of claim 65, wherein the composition further comprises proteins from serogroup B of N. meningitidis.
71:
- The composition of claim 66, wherein the composition further comprises proteins from serogroup B of N. meningitidis.
72:
- The composition of claim 68, wherein the composition further comprises proteins from serogroup B of N. meningitidis.
74:
- The composition of claim 73, wherein each conjugate comprises capsular saccharides that are oligosaccharides with an average degree of polymerization of less than 30 conjugated to the carrier protein selected from bacterial toxins and toxoids.
75:
- The composition of claim 74, wherein the bacterial toxins and toxoids are diphtheria toxoids or tetanus toxoids
76:
- The composition of claim 73, wherein each conjugate comprises a linker.
77:
- The composition of claim 73, wherein the capsular saccharide in each conjugate was activated or functionalized prior to conjugation.
78:
- The composition of claim 77, wherein the activation was with a cyanylating reagent.
79:
- The composition of claim 73, wherein the composition comprises MenA capsular saccharides, MenC capsular saccharides, MenW135 capsular saccharides, and MenY capsular saccharides.
80:
- The composition of claim 73, wherein the composition further comprises proteins from serogroup B of N. meningitidis.
81:
- The composition of claim 73, wherein the composition further comprises antigens from pneumococcus, hepatitis A virus, hepatitis B virus, B. pertussis, diphtheria, tetanus, Helicobacter pylori, polio and/or H. influenzae.
82:
- The composition of claim 73, wherein the composition is lyophilized.
83:
- A process for conjugating a bacterial capsular polysaccharide to a carrier protein, comprising:
- purifying the polysaccharide, comprising the steps of (a) precipitation of the polysaccharide using one or more cationic detergents, followed by (b) solubilisation of the precipitated polysaccharide using an alcohol,
- conjugating the polysaccharide to a carrier protein, wherein the carrier protein is a bacterial toxin or toxoid, and wherein conjugation is with a linker, and
- mixing the conjugated polysaccharide with a second bacterial capsular saccharide conjugated to a second carrier protein, wherein the second carrier protein is the bacterial toxin or toxoid,
- wherein the bacterial capsular polysaccharide is from Neisseria meningitidis serogroup A, W135 or Y, or from Haemophilus influenzae, or from Streptococcus pneumoniae.