Patent
The invention relates to activated Streptococcus pneumoniae serotype 10A, 22F or 33F polysaccharides and processes for their preparation. The invention also relates to immunogenic conjugates comprising Streptococcus pneumoniae serotype 10A, 22F or 33F polysaccharides covalently linked to a carrier protein, processes for their preparation and immunogenic compositions and vaccines comprising them.
1. A process for preparing an activated Streptococcus pneumoniae serotype 10A, 22F or 33F capsular polysaccharide, the process comprising the steps of:
(a) reducing the size of a purified serotype 10A, 22F or 33F capsular polysaccharide using high pressure homogenization to yield an isolated serotype 10A, 22F or 33F capsular polysaccharide;
(b) reacting the isolated serotype 10A, 22F or 33F capsular polysaccharide with an oxidizing agent; and
(c) quenching the oxidation reaction of step (b) by adding a quenching agent to produce an activated Streptococcus pneumoniae serotype 10A, 22F or 33F polysaccharide.
2. A process according to claim 1 wherein the concentration of serotype 10A, 22F or 33F polysaccharide in step (a) is between 0.5 and 5 mg/mL.
3. A process according to claim 1 wherein said capsular polysaccharide is reacted with 0.05 to 0.3 molar equivalents of oxidizing agent in step (b).
4. A process according to claim 1 wherein the oxidizing agent is periodate.
5. A process according to claim 1 wherein the quenching agent is selected from vicinal diols, 1,2-aminoalcohols, amino acids, glutathione, sulfite, bisulfate, dithionite, metabisulfite, thiosulfate, phosphites, hypophosphites or phosphorous acid.
6. A process according to claim 1 wherein the oxidation reaction is quenched by the addition of 0.1 to 10, 0.5 to 5, 0.5 to 3 or 0.5 to 2 molar equivalents of quenching agent.
7. A process according to claim 1 wherein said isolated capsular polysaccharide is an isolated Streptococcus pneumoniae serotype 10A capsular polysaccharide.
8. A process according to claim 1 wherein said isolated capsular polysaccharide is an isolated Streptococcus pneumoniae serotype 22F capsular polysaccharide.
9. A process according to claim 1 wherein said isolated capsular polysaccharide is an isolated Streptococcus pneumoniae serotype 33F capsular polysaccharide.