ACCELERATED METHOD OF PURIFICATION FOR OBTAINING CAPSULAR POLYSACCHARIDES OF STREPTOCOCCUS PNEUMONIAE Russian patent published in 2014 - IPC C12P19/04 C08B37/00 A61K39/09 

FIELD: chemistry.

SUBSTANCE: group of inventions relates to biotechnology. Claimed are versions of method of obtaining solution, which contains purified capsular polysaccharides, from lysate of cells of selected Streptococcus pneumoniae serotype. Lysis of cells of selected S. pneumoniae serotype is carried out. Obtained lysate is clarified by centrifugation or filtration. After that, ultrafiltration and diafiltration of clarified lysate are carried out in salt-free water with obtaining retentate. Retentate pH is reduced to the value lower than 4.5. Acidified retentate solution is kept to sediment precipitate. After that, retentate solution is filtered or centrifuged to obtain clarified solution of capsular polysaccharide. Then, clarified polysaccharide solution is filtered through filter with activated carbon. Obtained filtered solution id ultrafiltered and diafiltered. Obtained concentrated purified solution of capsular polysaccharide is filtered through sterile filter. S. pneumoniae serotypes are selected from group, consisting of serotypes 1, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F. Also claimed is modification of claimed method for serotype 19A. Said modification includes carrying out ultrafiltration and diafiltration before acidification at 4°C at pH 6 in sodium-phosphate buffer, keeping acidified retentate solution at for at least 2 hours 4°C and bringing pH of clarified polysaccharide solution to value 6 before the stage of filtration with active carbon. Also claimed are solutions, containing purified capsular polysaccharides of selected S. pneumoniae serotype, obtained by claimed methods, and their application in production of pneumococcal vaccine. Also claimed are purified capsular polysaccharides of S. pneumoniae serotype 6A with molecular weight 640000-670000 Da and 19A with molecular weight 488000-525000 Da, separated from said solutions.

EFFECT: accelerated method makes it possible to precipitate more than 98% and 90% of protein at the stages of acidification and adsorption respectively, without essential impact on polysaccharide output.

47 cl, 14 dwg, 19 tbl, 4 ex