Multivalent pneumococcal vaccines

A multivalent vaccine comprising twenty-four or more species of immunogenic complexes, wherein the twenty-four or more species of immunogenic complexes comprise:

• a first immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 1, and (b) a fusion protein;
• a second immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 2, and (b) a fusion protein;
• a third immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 3, and (b) a fusion protein;
• a fourth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 4, and (b) a fusion protein;
• a fifth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 5, and (b) a fusion protein;
• a sixth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 6A, and (b) a fusion protein;
• a seventh immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 6B, and (b) a fusion protein;
• an eighth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 7F, and (b) a fusion protein;
• a ninth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 8, and (b) a fusion protein;
• a tenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 9N, and (b) a fusion protein;
• an eleventh immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 9V, and (b) a fusion protein;
• a twelfth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 10A, and (b) a fusion protein;
• a thirteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 11A, and (b) a fusion protein;
• a fourteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 12F, and (b) a fusion protein;
• a fifteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 14, and (b) a fusion protein;
• a sixteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 15B, and (b) a fusion protein;
• a seventeenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 17F, and (b) a fusion protein;
• an eighteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 18C, and (b) a fusion protein;
• a nineteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 19A, and (b) a fusion protein;
• a twentieth immunogenic complex comprising (a) a biotinylated polysaccharide antigen, comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 19F, and (b) a fusion protein;
• a twenty-first immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 20B, and (b) a fusion protein;
• a twenty-second immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 22F, and (b) a fusion protein;
• a twenty-third immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 23F and (b) a fusion protein;
• and
• a twenty-fourth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 33F, and (b) a fusion protein;
• wherein each fusion protein comprises:
• (i) a biotin-binding moiety;
• (ii) a first polypeptide antigen comprising an SP1500 polypeptide or an antigenic fragment thereof;
• and
• (iii) a second polypeptide antigen comprising an SP0785 polypeptide or an antigenic fragment thereof;
• wherein each fusion protein comprises an amino acid sequence at least 95% identical to SEQ ID NO:6;
• and
• wherein in each species of the immunogenic complexes, the biotinylated polysaccharide antigen is non-covalently associated with the biotin-binding moiety of the fusion protein.

A method of making a multivalent vaccine, the method comprising a step of:

• mixing twenty-four or more species of immunogenic complexes in a single formulation, wherein the twenty-four or more species of immunogenic complexes comprise:
• a first immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 1, and (b) a fusion protein;
• a second immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 2, and (b) a fusion protein;
• a third immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 3, and (b) a fusion protein;
• a fourth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 4, and (b) a fusion protein;
• a fifth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 5, and (b) a fusion protein;
• a sixth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 6A, and (b) a fusion protein;
• a seventh immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 6B, and (b) a fusion protein;
• an eighth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 7F, and (b) a fusion protein;
• a ninth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 8, and (b) a fusion protein;
• a tenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 9N, and (b) a fusion protein;
• an eleventh immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 9V, and (b) a fusion protein;
• a twelfth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 10A, and (b) a fusion protein;
• a thirteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 11A, and (b) a fusion protein;
• a fourteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 12F, and (b) a fusion protein;
• a fifteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 14, and (b) a fusion protein;
• a sixteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 15B, and (b) a fusion protein;
• a seventeenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 17F, and (b) a fusion protein;
• an eighteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 18C, and (b) a fusion protein;
• a nineteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 19A, and (b) a fusion protein;
• a twentieth immunogenic complex comprising (a) a biotinylated polysaccharide antigen, comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 19F, and (b) a fusion protein;
• a twenty-first immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 20B, and (b) a fusion protein;
• a twenty-second immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 22F, and (b) a fusion protein;
• a twenty-third immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 23F and (b) a fusion protein;
• and
• a twenty-fourth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 33F, and (b) a fusion protein;
• wherein each fusion protein comprises:
• (i) a biotin-binding moiety;
• (ii) a first polypeptide antigen comprising an SP1500 polypeptide or an antigenic fragment thereof;
• and
• (iii) a second polypeptide antigen comprising an SP0785 polypeptide or an antigenic fragment thereof;
• wherein each fusion protein comprises an amino acid sequence at least 95% identical to SEQ ID NO:6;
• and
• wherein in each species of the immunogenic complexes, the biotinylated polysaccharide antigen is non-covalently associated with the biotin-binding moiety of the fusion protein.

A method of immunizing a subject against Streptococcus pneumoniae infection and/or colonization comprising administering to the subject a multivalent vaccine comprising twenty-four or more species of immunogenic complexes, wherein the twenty-four or more species of immunogenic complexes comprise:

• a first immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 1, and (b) a fusion protein;
• a second immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 2, and (b) a fusion protein;
• a third immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 3, and (b) a fusion protein;
• a fourth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 4, and (b) a fusion protein;
• a fifth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 5, and (b) a fusion protein;
• a sixth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 6A, and (b) a fusion protein;
• a seventh immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 6B, and (b) a fusion protein;
• an eighth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 7F, and (b) a fusion protein;
• a ninth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 8, and (b) a fusion protein;
• a tenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 9N, and (b) a fusion protein;
• an eleventh immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 9V, and (b) a fusion protein;
• a twelfth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 10A, and (b) a fusion protein;
• a thirteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 11A, and (b) a fusion protein;
• a fourteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 12F, and (b) a fusion protein;
• a fifteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 14, and (b) a fusion protein;
• a sixteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 15B, and (b) a fusion protein;
• a seventeenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 17F, and (b) a fusion protein;
• an eighteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 18C, and (b) a fusion protein;
• a nineteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 19A, and (b) a fusion protein;
• a twentieth immunogenic complex comprising (a) a biotinylated polysaccharide antigen, comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 19F, and (b) a fusion protein;
• a twenty-first immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 20B, and (b) a fusion protein;
• a twenty-second immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 22F, and (b) a fusion protein;
• a twenty-third immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 23F and (b) a fusion protein;
• and
• a twenty-fourth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen of Streptococcus pneumoniae serotype 33F, and (b) a fusion protein;
• wherein each fusion protein comprises:
• (i) a biotin-binding moiety;
• (ii) a first polypeptide antigen comprising an SP1500 polypeptide or an antigenic fragment thereof;
• and
• (iii) a second polypeptide antigen comprising an SP0785 polypeptide or an antigenic fragment thereof;
• wherein each fusion protein comprises an amino acid sequence at least 95% identical to SEQ ID NO:6;
• and
• wherein in each species of the immunogenic complexes, the biotinylated polysaccharide antigen is non-covalently associated with the biotin-binding moiety of the fusion protein.