Expression of pneumococcal surface protein a (pspa)

Abstract

The present invention relates to expression of Pneumococcal Surface Protein A (PspA). The invention represents an advancement in the field of genetic engineering and vaccine technology. The invention discloses expression vectors and recombinant host cells for expression of truncated PspA peptide. The invention also discloses vaccine compositions comprising the truncated peptides as carrier protein.

I/We claim: 
     
         1 . A pneumococcal conjugate vaccine comprising at least one capsular polysaccharide from  Streptococcus pneumoniae  serotypes selected from the group consisting of 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19F, 19A, 20A, 20B, 22F, 23A, 23B, 23F, 24B, 24F, 31, 33F, 34, 35B, 35F, 38, 39 and 45, wherein the polysaccharide is conjugated with a truncated PspA1 having a sequence of SEQ ID NO: 3 or SEQ ID NO: 4, or a combination of truncated PspA1 and at least one other carrier protein selected from group of carrier proteins comprising of CRM197, tetanus toxoid, pertussis toxoid, and PsaA. 
     
     
         2 . The pneumococcal conjugate vaccine of  claim 1 , wherein the at least one capsular polysaccharide from  Streptococcus pneumoniae  serotypes is selected from the group consisting of 3, 6A and 6B. 
     
     
         3 . The pneumococcal conjugate vaccine of  claim 1 , wherein the vaccine is a multivalent vaccine selected from 10 valent, 14 valent, 15 valent, 17 valent, 18 valent, 19 valent, 20 valent, 22 valent, 23 valent, 24 valent and 25 valent. 
     
     
         4 . The pneumococcal conjugate vaccine of  claim 1 , wherein the multivalent vaccine is a 15 valent composition comprising pneumococcal capsular polysaccharides selected from serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F. 
     
     
         5 . The pneumococcal conjugate vaccine of  claim 1  comprising capsular polysaccharides from at least three  Streptococcus pneumoniae  serotypes 3, 6A and 6B conjugated to truncated PspA1 as set forth in SEQ ID NO: 3 or SEQ ID NO: 4, and capsular polysaccharides from  Streptococcus pneumoniae  serotypes 1, 4, 5, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F conjugated to CRM197. 
     
     
         6 . The pneumococcal conjugate vaccine of  claim 5 , wherein the composition comprises:
 2.2 to 4.4 μg of each of the capsular polysaccharides from  Streptococcus pneumoniae  serotypes 3, 6A and 6B conjugated to truncated PspA1; and   2.2 μg of each of the capsular polysaccharides from  Streptococcus pneumoniae  serotypes 1, 4, 5, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F conjugated to CRM197.   
     
     
         7 . The pneumococcal conjugate vaccine of  claim 1  wherein the pneumococcal vaccine composition is a single 0.5 mL dose comprising:
 about 2.2 to 4.4 μg of one or more pneumococcal capsular polysaccharides; 
 about 1 μg to about 30 μg of truncated PspA1, having a sequence of SEQ ID NO: 3 or SEQ ID NO: 4, conjugated to each of the one or more pneumococcal capsular polysaccharides; 
 about 0.2 mg to about 1 mg of aluminum phosphate adjuvant; and 
 an excipient. 
 
     
     
         8 . The pneumococcal conjugate vaccine of  claim 7 , wherein the composition further comprises about 1 μg to about 30 μg of CRM 197  conjugated to each of the one or more pneumococcal capsular polysaccharides. 
     
     
         9 . The pneumococcal conjugate vaccine of  claim 1 , wherein truncated PspA1 is a carrier protein in a conjugate vaccine. 
     
     
         10 . The pneumococcal conjugate vaccine of  claim 1 , wherein the pneumococcal capsular polysaccharide is conjugated to the truncated PspA1 by a method comprising:
 a. preparing the truncated PspA1;   b. activating the capsular polysaccharide from one or more pneumococcal serotypes to obtain an activated polysaccharide; and   c. conjugating the activated polysaccharide with the truncated PspA1 prepared in step (a) to obtain a pneumococcal capsular polysaccharide-carrier protein conjugate.   
     
     
         11 . The pneumococcal conjugate vaccine of  claim 10 , wherein the truncated PspA1 is reacted with the activated polysaccharide in a ratio of 1:1. 
     
     
         12 . A method of conjugating a pneumococcal capsular polysaccharide to a truncated PspA1 having a sequence of SEQ ID NO: 3 or SEQ ID NO: 4 comprising:
 a. preparing the truncated PspA1;   b. activating the capsular polysaccharide from one or more pneumococcal serotypes to obtain an activated polysaccharide; and   c. conjugating the activated polysaccharide with the truncated PspA1 prepared in step (a) to obtain a pneumococcal capsular polysaccharide-carrier protein conjugate.   
     
     
         13 . The method of  claim 12 , wherein the truncated PspA1 is reacted with the activated polysaccharide in a ratio of 1:1. 
     
     
         14 . The method of  claim 12 , wherein the pneumococcal capsular polysaccharide comprises one or more  Streptococcus pneumoniae  serotypes selected from the group consisting of 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19F, 19A, 20A, 20B, 22F, 23A, 23B, 23F, 24B, 24F, 31, 33F, 34, 35B, 35F, 38, 39 and 45. 
     
     
         15 . The method of  claim 14 , wherein the capsular polysaccharide comprises one or more  Streptococcus pneumoniae  serotypes selected from the group consisting of 3, 6A and 6B. 
     
     
         16 . The method of  claim 14  wherein the capsular polysaccharide comprises a  Streptococcus pneumoniae  serotypes selected from the group consisting of 1, 4, 5, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F conjugated to CRM 197 . 
     
     
         17 . The method of  claim 16 , wherein the composition further comprises about 1 μg to about 30 μg of CRM 197  conjugated to each of the one or more pneumococcal capsular polysaccharides. 
     
     
         18 . The method of  claim 12 , wherein truncated PspA1 is a carrier protein in a conjugate vaccine. 
     
     
         19 . The method of  claim 12 , wherein the capsular polysaccharide is further conjugated to at least one other carrier protein selected from group of carrier proteins comprising of CRM197, tetanus toxoid, pertussis toxoid, and PsaA. 
     
     
         20 . The method of  claim 12 , wherein the pneumococcal capsular polysaccharide-carrier protein conjugate is in a conjugate vaccine.