IMMUNOGENIC EBOLAVIRUS FUSION PROTEINS AND RELATED METHODS
Provided are fusion proteins including an artificially modified amino acid sequence of an ebolavirus glycoprotein (GP) joined to an amino acid sequence of a ferritin subunit polypeptide. An artificially modified amino acid sequence of an ebolavirus GP includes artificially installed glycosylation sites. Nanoparticles including fusion proteins, with surface-exposed trimers of the ebolavirus GP, are also provided. Also provided are nucleic acids and vectors encoding the fusion proteins, cells containing such nucleic acid and vectors, immunogenic compositions including the fusion proteins, the nanoparticles, or the vectors, as well as corresponding methods and kits.
1 . A fusion protein of an artificially modified amino acid sequence of an ebolavirus glycoprotein (GP) and an amino acid sequence of a ferritin subunit polypeptide, wherein the artificially modified amino acid sequence of the ebolavirus GP is a sequence with at least 90% sequence identity to SEQ ID NO:13, SEQ ID NO:14, or SEQ ID NO: 15.
2 . (canceled)
3 . The fusion protein of claim 1 , wherein the artificially modified amino acid sequence of the ebolavirus GP comprises a deletion of the mucin-like domain.
4 . The fusion protein of claim 1 , wherein the artificially modified amino acid sequence of the ebolavirus GP comprises at least two artificial glycosylation sites.
5 . (canceled)
6 . The fusion protein of claim 4 , wherein the artificial glycosylation sites are located in the glycan cap domain of the ebolavirus GP.
7 . The fusion protein of claim 4 , wherein the artificially modified amino acid sequence of the ebolavirus GP contains two or more amino acid substitutions at positions corresponding to positions 251, 280, 272, 274, 212, and 214 of SEQ ID NO:12, wherein the two or more amino acid substitutions result in the artificial glycosylation sites.
8 . The fusion protein of claim 7 , wherein the two or more amino acid substitutions are D251N, L280N, S272N, E274T, Q212N, and E212T.
9 - 14 . (canceled)
15 . The fusion protein of claim 1 , wherein the amino acid sequence of the fusion protein is a sequence with at least 90% sequence identity to SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:1.
16 . A nanoparticle comprising an oligomer of the fusion protein of claim 1 .
17 - 18 . (canceled)
19 . A nucleic acid encoding the fusion protein of claim 1 .
20 . The nucleic acid of claim 19 , wherein the nucleic acid is DNA or RNA.
21 . A vector comprising the nucleic acid of claim 19 .
22 . A cell comprising the nucleic acid of claim 19 .
23 . The cell of claim 22 , wherein the cell is a mammalian cell.
24 - 26 . (canceled)
27 . A cell culture comprising a plurality of cells of claim 22 .
28 . An immunogenic composition comprising the fusion protein of claim 1 .
29 . An immunogenic composition comprising two or more different fusion proteins of claim 1 .
30 - 31 . (canceled)
32 . A kit comprising the immunogenic composition of claim 28 and one or more of: a device for administering the immunogenic composition, and an excipient.
33 . A method of inducing an immune response in a subject, the method comprising administering to the subject the immunogenic composition of claim 28 .
34 - 37 . (canceled)
38 . A method of producing the fusion protein, comprising:
introducing into a cell the nucleic acid of claim 19 ;
incubating the cell under conditions allowing for expression of the fusion protein; and
isolating the fusion protein.
39 - 40 . (canceled)
41 . A method of producing a nanoparticle, comprising:
introducing into a cell the nucleic acid of claim 19 ;
incubating the cell under conditions allowing for expression of the fusion protein and self-assembly of the nanoparticle; and
isolating the nanoparticle.
42 - 43 . (canceled)