Multi-functionalized nOMV conjugates

An immunogenic conjugate comprising an isolated, intact native outer membrane vesicle (nOMV), having at least a surface saccharide moiety connected to an antigen, and having at least a surface protein residue connected to a different antigen through a bivalent linker;

• wherein said bivalent linker has the general formula (I):
• X-L-X′ (I)
• wherein:
• X and X′ are different to each other or the same, and are a functional group able to selectively react with the nOMV protein residue on one hand and with the antigen on the other hand;
• -L- is a bivalent linear or branched C1-C15 alkyl or alkenyl group optionally substituted, and optionally interrupted by one or more heteroatom selected from:
• oxygen (—O—), sulfur (—S—), and nitrogen (NH— or optionally substituted —N— group);
• wherein X and X′ are N-hydroxysuccinimide ester derivatives selected from at least one of:
• wherein the * represents the point of contact with the -L- group in formula (I);
• wherein said nOMV is prepared from S. sonnei, S. flexneri, meningococcus, or Salmonella bacteria;
• wherein said antigen is selected from a group consisting of:
• Neisseria meningitidis fHbp, SsIE, and FdeC;
• wherein SsIE comprise an amino acid sequence having 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity to SEQ ID NO:
• 15;
• wherein FdeC comprise an amino acid sequence having 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity to SEQ ID NO:
• 16;
• and
• wherein Neisseria meningitidis fHbp comprise an amino acid sequence comprising SEQ ID NO:
• 8, SEQ ID NO:
• 1, or of SEQ ID NO:
• 9.