Multivalent pneumococcal vaccines

A method of immunizing a subject against Streptococcus pneumoniae infection and/or colonization comprising administering to the subject an immunologically effective amount of a vaccine, wherein the vaccine comprises an immunogenic complex that comprises:

• (a) a biotinylated polysaccharide;
• and
• (b) a fusion protein comprising an amino acid sequence at least 90% identical to SEQ ID NO:
• 19, SEQ ID NO:
• 20, SEQ ID NO:
• 58, or SEQ ID NO:
• 59, wherein the fusion protein comprises:
• (i) a biotin-binding moiety;
• (ii) a non-hemolytic pneumolysin (Ply) polypeptide comprising mutations at amino acid residues corresponding to amino acid residues 294, 385, 428, and 433 of wild-type Streptococcus pneumoniae pneumolysin (SEQ ID NO:
• 3) or antigenic fragment thereof;
• and
• (iii) an SP0435 polypeptide or antigenic fragment thereof;
• wherein the biotinylated polysaccharide antigen is non-covalently associated with the biotin-binding moiety of the fusion protein to form the immunogenic complex.