Patent ES-8602417-A1 (ES)
Title: Multivalent pneumococcal vaccine and preparation thereof.
IPC: C12N/
Filed: 1984-06-08
Abstract: Multivalent pneumococcal vaccine comprises immunologically active purified pneumococcal capsular polysaccharide (free from C polysaccharide) of Types 1-5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F, opt. with 6A and/or 25. - The Type 3 polysaccharide is obtd. in purified form from a clarified fermentation lysate by (a) pptn. with 0.15-0.5 vol. of alcohol, the lysate being at about pH 6.7 and having a final Na acetate concn. of about 4% and being at 2-6 deg.C (b) redissolving the ppte. (c) repeating step (a) with 0.25-0.6 vol. alcohol, then step (b) (d) repeating step (b) with 0.1-0.5 vol. alcohol, then step (b) (e) fractional pptn. of impurities from the prod. with hexadecyltrimethylammonium bromide (I) at 21-5 deg.C and pH 7.4+-0.1 at a (I) concn. of 0.1-0.5 vol.%, based on a 10% (I) soln. (f) repptn. of the polysaccharide at about pH 6.7 with 4% Na acetate and about 0.5 vol. alcohol, and redissolving the ppte. in pyrogen-free water (g) repeating step (f) twice (h) purifying the polysaccharide soln. with activated charcoal at pH 6.1 and a NaCl concn. of 0.15M by adding the charcoal in suspn. to 3-7% based on a 20% activated charcoal suspn., chilling the soln. for 30 mins. and filtering (j) diafiltering the soln. against distd. water and (k) freezing and lyophilising the prod. (0/0) DEAB- DE3485778 G Multivalent pneumococcal vaccine comprises immunologically active purified pneumococcal capsular polysaccharide (free from C polysaccharide) of Types 1-5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F, opt. with 6A and/or 25. - The Type 3 polysaccharide is obtd. in purified form from a clarified fermentation lysate by (a) pptn. with 0.15-0.5 vol. of alcohol, the lysate being at about pH 6.7 and having a final Na acetate concn. of about 4% and being at 2-6 deg.C (b) redissolving the ppte. (c) repeating step (a) with 0.25-0.6 vol. alcohol, then step (b) (d) repeating step (b) with 0.1-0.5 vol. alcohol, then step (b) (e) fractional pptn. of impurities from the prod. with hexadecyltrimethylammonium bromide (I) at 21-5 deg.C and pH 7.4+-0.1 at a (I) concn. of 0.1-0.5 vol.%, based on a 10% (I) soln. (f) repptn. of the polysaccharide at about pH 6.7 with 4% Na acetate and about 0.5 vol. alcohol, and redissolving the ppte. in pyrogen-free water (g) repeating step (f) twice (h) purifying the polysaccharide soln. with activated charcoal at pH 6.1 and a NaCl concn. of 0.15M by adding the charcoal in suspn. to 3-7% based on a 20% activated charcoal suspn., chilling the soln. for 30 mins. and filtering (j) diafiltering the soln. against distd. water and (k) freezing and lyophilising the prod. EPAB- EP-157899 B Multivalent pneumococcal vaccine comprises immunologically active purified pneumococcal capsular polysaccharide (free from C polysaccharide) of Types 1-5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F, opt. with 6A and/or 25. - The Type 3 polysaccharide is obtd. in purified form from a clarified fermentation lysate by (a) pptn. with 0.15-0.5 vol. of alcohol, the lysate being at about pH 6.7 and having a final Na acetate concn. of about 4% and being at 2-6 deg.C (b) redissolving the ppte. (c) repeating step (a) with 0.25-0.6 vol. alcohol, then step (b) (d) repeating step (b) with 0.1-0.5 vol. alcohol, then step (b) (e) fractional pptn. of impurities from the prod. with hexadecyltrimethylammonium bromide (I) at 21-5 deg.C and pH 7.4+-0.1 at a (I) concn. of 0.1-0.5 vol.%, based on a 10% (I) soln. (f) repptn. of the polysaccharide at about pH 6.7 with 4% Na acetate and about 0.5 vol. alcohol, and redissolving the ppte. in pyrogen-free water (g) repeating step (f) twice (h) purifying the polysaccharide soln. with activated charcoal at pH 6.1 and a NaCl concn. of 0.15M by adding the charcoal in suspn. to 3-7% based on a 20% activated charcoal suspn., chilling the soln. for 30 mins. and filtering (j) diafiltering the soln. against distd. water and (k) freezing and lyophilising the prod. (63pp Dwg.No.0/0) - EP-157899 B A multivalent pneumococcal vaccine comprising effective amounts of immunologically active purified pneumococcal capsular polysaccharide (substantially absent ''C'' polysaccharide) of pneumococcal types (Danish designation) consisting of 1,2,3,4,5,6B,7F,8,9N,10A,11A,12F,1 4,15B,17F,18C,19A,19F,20,22F,23- F,33F, and from none to one or both of 6A and 25 wherein effectively immunogenic Type 3 pneumococcal polysaccharide is purified from a clarified fermentation lysate by the process of (a) precipitating with 0.15 and 0.5 volumes of alcohol, the lysate being at pH of about 6.7 and having a final sodium acetate concentration of about 4% and being at a temperature of 2-6 deg.C, and (b) redissolving such precipitate, and (c) repeating Step (a) at about 0.25 to 0.6 volumes of alcohol and (d) repeating step (b), and (e) repeating Step (a) with 0.1 to 0.5 volumes of alcohol, and (f) repeating Step (b), and (g) fractionally precipitating impurities from the polysaccharide of (f) with hexadecyltrimethylammonium bromide at a temperature of from 21-25 deg.C and a pH of 7.4+/-0.1, and a hexadecyltrimethylammonium bromide concentration of 0.1 to 0.5 volumes percent based on a 10% hexadecyltrimethylammonium bromide solution, and (h) reprecipitating the polysacharide at a pH of about 6.7 with about 4% sodium acetate and about a 0.50 volume of alcohol, and redissolving in pyrogen-free water, and (i) repeating Step (h) 2 times, and (j) purifying the polysaccharide solution of (i) with activated charcoal at a pH of 6.1 and a sodium chloride concentration of 0.15M, by adding activated charcoal in suspension to a concentration of 3% to 7% based on a 20% activated charcoal suspension, and the solution standing chilled for about 30 minutes and filtering out said activated charcoal, and (k) diafiltering the solution against distilled water, and (l) freezing and lyophilizing the resultant product.p - EP-157899 B A multivalent pneumococcal vaccine comprising effective amounts of immunologically active purified pneumococcal capsular polysaccharide (substantially absent "C" polysaccharide) of pneumococcal types (Danish designation) consisting of 1,2,3,4,5,6B,7F,8,9N,10A,11A,12F,1 4,15B,17F,18C,19A,19F,20,22F,23F,33F, and from none to one or both of 6A and 25 wherein effectively immunogenic Type 3 pneumococcal polysaccharide is purified from a clarified fermentation lysate by the process of (a) precipitating with 0.15 and 0.5 volumes of alcohol, the lysate being at pH of about 6.7 and having a final sodium acetate concentration of about 4% and being at a temperature of 2-6 deg.C, and (b) redissolving such precipitate, and (c) repeating Step (a) at about 0.25 to 0.6 volumes of alcohol and (d) repeating step (b), and (e) repeating Step (a) with 0.1 to 0.5 volumes of alcohol, and (f) repeating Step (b), and (g) fractionally precipitating impurities from the polysaccharide of (f) with hexadecyltrimethylammonium bromide at a temperature of from 21-25 deg.C and a pH of 7.4+/-0.1, and a hexadecyltrimethylammonium bromide concentration of 0.1 to 0.5 volumes percent based on a 10% hexadecyltrimethylammonium bromide solution, and (h) reprecipitating the polysaccharide at a pH of about 6.7 with about 4% sodium acetate and about a 0.50 volume of alcohol, and redissolving in pyrogen-free water, and (i) repeating Step (h) 2 times, and (j) purifying the polysaccharide solution of (i) with activated charcoal at a pH of 6.1 and a sodium chloride concentration of 0.15M, by adding activated charcoal in suspension to a concentration of 3% to 7% based on a 20% activated charcoal suspension, and the solution standing chilled for about 30 minutes and filtering out said activated charcoal, and (k) diafiltering the solution against distilled water, and (l) freezing and lyophilizing the resultant product. USAB- US4686102 A Effective immunogenic type 3 pneumococcal polysaccharide is purified from a clarified fermentation lysate by (A) pptg. the lysate with 0.15-0.5 vol. alcohol at pH 6.7 and a final Na acetate concn. 4% and being at a temp. 2-6 deg.C, (B) redissolving the ppte. and repeating step (A) at 0.25-0.6 vol. alcohol, (C) repeating step (B) and repeating step (A) with 0.1-0.5 vol. alcohol, (D) repeating step (B), fractionally pptg. impurities from the polysaccharide with hexadecyl-Me3NBr at 21-25 deg.C at pH 7.3-7.5 and a bromide concn. of 0.1-0.5 vol.% referred to a 10% bromide soln. (E) repptg. the polysaccharide at pH 6.7 and 4% Na acetate and 0.50 vol. alcohol and redissolving it in pyrogen free water, (F) repeating step (E) twice and purifying the polysaccharide soln. obtd. with activated charcoal at pH 6.1 and an NaCl concn. 0.15 M by adding a 20% suspension of activated charcoal to a concn. of 3-7%, chilling the soln. and filtering off the activated charcoal after 30 min. standing, and (G) diafiltering the soln. against distilled water and freezing and lyophilising the prod. obtd.