Theragnostic particles
The present invention provides viral-based nanoparticles for therapeutic and diagnostic use, and methods for making and using the nanoparticles. Specifically, such nanoparticles comprise decoration-competent viral particles shells such as expanded capsids of phages, stabilized with engineered decoration proteins that have been linked to one or more compounds not naturally occurring on a wild type viral capsid.
1. A theragnostic particle enveloped by a viral particle shell, wherein:
at least one engineered decoration protein is bound to the outer surface of the shell,
the at least one engineered decoration protein comprises the amino acid sequence of SEQ ID NO: 4 (wt gpD), wherein at least one solvent-exposed residue of the at least one engineered decoration protein is mutated to a cysteine or a non-natural amino acid, and
the at least one solvent-exposed residue is selected from the group consisting of Ser29, Ala30, Lys31, Asp41, Thr42, Ser43, Ser44, Arg45, Lys46, Gly52, Thr53, Thr54, Asp55, Asp67, Gln68, Thr69, Ser70, Thr71, Thr72, Glu90, Ala9, Ala92, Ser93, Asp94, Glu95, Thr96, Lys97, Lys98, Arg99, and Thr100 as corresponding to the amino acid sequence of SEQ ID NO: 4.
2. The particle of claim 1 , wherein the shell comprises at least one decoration competent viral particle shell selected from the group consisting of bacteriophage lambda, bacteriophage T4, bacteriophage L, bacteriophage P22, bacteriophage 21, bacteriophage P4, herpesvirus, and adenovirus.
3. The particle of claim 1 , wherein the shell comprises a bacteriophage lambda viral particle shell.
4. The particle of claim 1 , wherein the shell comprises an expanded viral particle shell.
5. The particle of claim 1 , wherein the at least one mutated residue is independently conjugated with one or more compounds independently selected from the group consisting of a proteinaceous agent and a non-proteinaceous agent.
6. The particle of claim 4 , wherein the one or more compounds do not naturally occur a in a wild type viral capsid.
7. The particle of claim 4 , wherein the one or more compounds are present on the particle in a defined ratio relative to the engineered decoration protein.
8. The particle of claim 4 , wherein the one or more compounds comprise two or more different compounds, which are present on the particle in a defined ratio relative to each other.
9. The particle of claim 4 , wherein the non-proteinaceous agent is selected from the group consisting of nucleic acids, lipids, carbohydrates, polypeptides, polymers, organic molecules, inorganic molecules, and any combinations thereof.
10. The particle of claim 4 , wherein the non-proteinaceous agent is selected from the group consisting of therapeutic compounds, diagnostic compounds, adjuvants, antigens, antibodies, and any combinations thereof.
11. A pharmaceutical composition comprising the theragnostic particle of claim 1 and a pharmaceutically acceptable carrier.
12. An isolated recombinant protein comprising the amino acid sequence of SEQ ID NO: 4 (wt gpD), wherein at least one residue of the protein is mutated to a cysteine or a non-natural amino acid, wherein the at least one residue is selected from the group consisting of Ser29, Ala30, Lys1, Asp41, Thr42, Ser43, Ser44, Arg45, Lys46, Gly52, Thr53, Thr54, Asp55, Asp67, Gln68, Thr69, Ser70, Thr71, Thr72, Glu90, Ala91, Ala92, Ser93, Asp94, Glu95, Thr96, Lys97, Lys98, Arg99, and Thr100 as corresponding to the amino acid sequence of SEQ ID NO: 4.
13. The protein of claim 12 , wherein the at least one mutated residue is independently conjugated with one or more compounds independently selected from the group consisting of a proteinaceous agent and a non-proteinaceous agent.
14. An isolated nucleic acid encoding the recombinant protein of claim 12 .
15. A recombinant expression vector comprising the isolated nucleic acid of claim 14 .
16. An isolated host cell comprising the recombinant expression vector of claim 15 .
17. An in vitro method for preparing a theragnostic particle of claim 1 , the method comprising decorating a decoration competent viral particle shell in vitro with a defined amount of at least one engineered decoration protein linked to one or more compounds, wherein the at least one engineered decoration protein stabilizes the decoration competent viral particle shell to produce a theragnostic particle, wherein the at least one engineered decoration protein is as recited in claim 1 .
18. The method of claim 17 , wherein the decoration competent viral particle shell is an expanded viral particle shell, and wherein method comprises contacting the viral particle with an expansion agent, thus generating the expanded viral particle shell.
19. The method of claim 18 , wherein the expansion agent is selected from the group consisting of chaotropic agents, pH, heat, and any combinations thereof.
20. The method of claim 17 , wherein the decoration competent viral particle shell is at least one selected from the group consisting of a bacteriophage lambda, bacteriophage T4, bacteriophage L, bacteriophage P22, bacteriophage 21, bacteriophage L, bacteriophage P4, herpesvirus, and adenovirus.