Multivalent nanoparticle-based vaccines
Novel, nanoparticle-based vaccines are provided that elicit an immune response to a broad range of infectious agents, such as influenza viruses. The nanoparticles comprise a heterogeneous population of fusion proteins, each comprising a monomeric subunit of a self-assembly protein, such as ferritin, joined to one or more immunogenic portions of a protein from an infectious agent, such as influenza virus. The fusion proteins self-assemble to form nanoparticles that display a heterogeneous population of immunogenic portions on their surface. When administered to an individual, such nanoparticles elicit an immune response to different strains, types, subtypes and species within the same taxonomic family. Thus, such nanoparticles can be used to vaccinate an individual against infection by different Types, subtypes and/or strains of infectious agents. Also provided are specific fusion proteins, nucleic acid molecules encoding such fusion proteins and methods of using nanoparticles of the invention to vaccinate individuals.
1 . A method of producing a nanoparticle, comprising:
culturing a cell comprising at least two nucleic acid molecules, wherein:
each nucleic acid molecule encodes a unique species of self-assembling fusion protein, each fusion protein comprises a self-assembling, monomeric subunit protein joined to an immunogenic portion of an influenza virus hemagglutinin (HA) protein, wherein the immunogenic portion of the influenza virus HA protein is a receptor binding domain (RBD);
the immunogenic portion in each species of self-assembling fusion protein differs from the immunogenic portion in the other species of self-assembling fusion proteins by at least one amino acid;
the fusion proteins self-assemble to form a nanoparticle that displays on its surface, the immunogenic portions of the at least two species of self-assembling fusion proteins;
at least one of the species of self-assembling fusion proteins comprises an amino acid sequence at least 90% identical to SEQ ID NO: 97; and
the cell is cultured under conditions suitable for expressing the encoded self-assembling fusion proteins to form the nanoparticle.
2 . The method of claim 1 , wherein at least one of the species of self-assembling fusion proteins comprises an amino acid sequence at least 95% identical to SEQ ID NO: 97.
3 . The method of claim 1 , wherein at least one of the species of self-assembling fusion proteins comprises an amino acid sequence at least 99% identical to SEQ ID NO: 97.
4 . The method of claim 1 , wherein the at least one species of self-assembling fusion protein comprises an amino acid sequence set forth as SEQ ID NO:97, SEQ ID NO: 100, SEQ ID NO:112, SEQ ID NO: 115, SEQ ID NO:118, SEQ ID NO:121, SEQ ID NO: 124, SEQ ID NO: 127, SEQ ID NO: 130, SEQ ID NO:133, SEQ ID NO:136, SEQ ID NO: 139, or SEQ ID NO:142.
5 . The method of claim 1 , wherein the at least one species of self-assembling fusion protein comprises an amino acid sequence set forth as SEQ ID NO: 97.
6 . A method of producing a nanoparticle, comprising:
culturing a cell comprising at least four nucleic acid molecules, wherein:
each nucleic acid molecule encodes a unique species of self-assembling fusion protein, each fusion protein comprises a self-assembling, monomeric subunit protein joined to an immunogenic portion of an influenza virus hemagglutinin (HA) protein, wherein the immunogenic portion of the influenza virus HA protein is a receptor binding domain (RBD);
the immunogenic portion in each species of self-assembling fusion protein differs from the immunogenic portion in the other species of self-assembling fusion proteins by at least one amino acid;
the fusion proteins self-assemble to form a nanoparticle that displays on its surface, the immunogenic portions of the at least four species of self-assembling fusion proteins;
at least one of the species of self-assembling fusion proteins comprises an amino acid sequence at least 90% identical to SEQ ID NO: 97; and
the cell is cultured under conditions suitable for expressing the encoded self-assembling fusion proteins to form the nanoparticle.
7 . The method of claim 6 , wherein at least one of the species of self-assembling fusion proteins comprises an amino acid sequence at least 95% identical to SEQ ID NO: 97.
8 . The method of claim 6 , wherein at least one of the species of self-assembling fusion proteins comprises an amino acid sequence at least 99% identical to SEQ ID NO: 97.
9 . The method of claim 6 , wherein the at least one species of self-assembling fusion protein comprises an amino acid sequence set forth as SEQ ID NO:97, SEQ ID NO:100, SEQ ID NO:112, SEQ ID NO: 115, SEQ ID NO:118, SEQ ID NO:121, SEQ ID NO: 124, SEQ ID NO:127, SEQ ID NO: 130, SEQ ID NO: 133, SEQ ID NO:136, SEQ ID NO:139, or SEQ ID NO:142.
10 . The method of claim 6 , wherein the at least one species of self-assembling fusion protein comprises an amino acid sequence set forth as SEQ ID NO: 97.