Self-assembling insect ferritin nanoparticles
Disclosed are recombinant insect ferritin nanoparticles that can be used to display two different trimeric antigens at an equal ratio. Also disclosed are nucleic acids encoding the recombinant insect ferritin nanoparticles and methods of producing the recombinant insect ferritin nanoparticles. Methods for eliciting an immune response in a subject are also provided.
1. A recombinant insect ferritin nanoparticle, comprising:
twelve recombinant ferritin heavy chains and twelve recombinant ferritin light chains self-assembled into a globular ferritin nanoparticle; wherein
(A)
the recombinant insect ferritin heavy chains consist of 172 to 174 amino acids from the C-terminus of an insect ferritin heavy chain;
the recombinant insect ferritin light chains consist of 182 to 184 amino acids or 177 amino acids from the C-terminus of an insect ferritin light chain; and
each of the ferritin heavy and light chains is fused N terminally to a viral protein; and
(B)
the recombinant ferritin heavy chains consist of an amino acid sequence at least 90% identical to SEQ ID NO: 2; and the recombinant ferritin light chains consist of an amino acid sequence at least 90% identical to SEQ ID NO: 6; or
the recombinant ferritin heavy chains each consist of an amino acid sequence at least 90% identical to SEQ ID NO: 4; and the recombinant ferritin light chains each consist of an amino acid sequence at least 90% identical to SEQ ID NO: 8.
2. The recombinant insect ferritin nanoparticle of claim 1 , wherein the recombinant ferritin heavy chains consist of an amino acid sequence at least 90% identical to SEQ ID NO: 2; and the recombinant ferritin light chains consist of an amino acid sequence at least 90% identical to SEQ ID NO: 6.
3. The recombinant insect ferritin nanoparticle of claim 1 , wherein the recombinant ferritin heavy chains consist of an amino acid sequence at least 95% identical to SEQ ID NO: 2; and the recombinant ferritin light chains consist of an amino acid sequence at least 95% identical to SEQ ID NO: 6.
4. The recombinant insect ferritin nanoparticle of claim 1 , wherein the recombinant ferritin heavy chains consist of the amino acid sequence set forth as SEQ ID NO: 2, and the recombinant ferritin light chains consist of the amino acid sequence set forth as SEQ ID NO: 6.
5. The recombinant insect ferritin nanoparticle of claim 1 , wherein the recombinant ferritin heavy chains each consist of an amino acid sequence at least 90% identical to SEQ ID NO: 4; and the recombinant ferritin light chains each consist of an amino acid sequence at least 90% identical to SEQ ID NO: 8.
6. The recombinant insect ferritin nanoparticle of claim 1 , wherein the recombinant ferritin heavy chains each consist of an amino acid sequence at least 95% identical to SEQ ID NO: 4; and the recombinant ferritin light chains each consist of an amino acid sequence at least 95% identical to SEQ ID NO: 8.
7. The recombinant insect ferritin nanoparticle of claim 1 , wherein the recombinant ferritin heavy chains consist of the amino acid sequence set forth as SEQ ID NO: 4, and the recombinant ferritin light chains consist of the amino acid sequence set forth as SEQ ID NO: 8.
8. The recombinant insect ferritin nanoparticle of claim 1 , wherein the viral protein is a viral envelope protein.
9. The recombinant insect ferritin nanoparticle of claim 8 , wherein the viral envelope protein is from any one of HIV-1, influenza, RSV, MPV, HPIV, Ebola, Marburg, MERS coronavirus, or SARS coronavirus.
10. The recombinant insect ferritin nanoparticle of claim 8 , wherein the viral envelope protein is an influenza HA stem protein.
11. An isolated nucleic acid molecule encoding the recombinant insect ferritin light chain and/or the recombinant insect ferritin heavy chain of claim 1 .
12. The nucleic acid molecule of claim 11 , operably linked to a promoter.
13. The nucleic acid molecule of claim 11 , wherein the nucleic acid molecule is DNA or mRNA.
14. An expression vector comprising the nucleic acid molecule of claim 11 .
15. An isolated host cell comprising the expression vector of claim 14 .
16. A method of producing a recombinant insect ferritin nanoparticle, comprising:
transfecting a permissive cell culture with the vector of claim 14 ;
incubating the cell culture for a sufficient period of time to allow for protein expression from the vector; and
purifying the insect ferritin nanoparticle.
17. A composition comprising the recombinant insect ferritin nanoparticle of claim 1 , and a pharmaceutically acceptable carrier.
18. A method of inducing an immune response to a viral envelope protein in a subject, comprising administering an effective amount of the recombinant insect ferritin nanoparticle of claim 8 , or a nucleic acid molecule encoding the recombinant insect ferritin light chain and the recombinant insect ferritin heavy chain of the recombinant nanoparticle, to the subject to induce the immune response.
19. The method of claim 18 , wherein the nucleic acid molecule is an mRNA molecule, and the recombinant insect ferritin light chain and the recombinant insect ferritin heavy chain of the recombinant nanoparticle are linked to influenza HA stem proteins.