Virus-like particle conjugates for diagnosis and treatment of tumors
The present disclosure is directed to methods and compositions for the diagnosis and/or treatment of tumors, such as ocular tumors, using virus-like particles conjugated to photosensitive molecules.
1. A method of producing tumor-targeting bioconjugates, the method comprising:
(a) transfecting cells in vitro with deoxyribonucleic acid (DNA) encoding papilloma virus capsid proteins, wherein expression of the DNA in the cells results in production of the papilloma virus capsid proteins, and the papilloma virus capsid proteins assemble to form proto-capsids;
(b) collecting the proto-capsids and subjecting the proto-capsids to a maturation process to form virus-like particles comprising the papilloma virus capsid proteins; and
(c) conjugating near infrared phthalocyanine dye molecules to papilloma virus capsid proteins of the virus-like particles, thereby producing the tumor-targeting bioconjugates, each comprising about 50 to about 1000 near infrared phthalocyanine dye molecules, wherein the near infrared phthalocyanine dye molecules become toxic or produce a toxic molecule upon light activation.
2. The method of claim 1 further comprising treating the papilloma virus capsid proteins of step (a) with benzonase to eliminate host cell DNA.
3. The method of claim 1 , wherein the papilloma virus capsid proteins are human papilloma virus (HPV) capsid proteins.
4. The method of claim 3 , wherein the HPV capsid proteins comprise HPV L1 capsid proteins or a combination of HPV L1 capsid proteins and HPV L2 capsid proteins.
5. The method of claim 4 , wherein the HPV L1 capsid proteins comprise an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 1.
6. The method of claim 1 , wherein the papilloma virus capsid proteins are non-human papilloma virus capsid proteins.
7. The method of claim 6 , wherein the non-human papilloma virus capsid proteins are bovine papilloma virus (BPV) capsid proteins.
8. The method of claim 7 , wherein the BPV capsid proteins comprise BPV L1 capsid proteins or a combination of BPV L1 capsid proteins and BPV L2 capsid proteins.
9. The method of claim 8 , wherein the BPV L1 capsid proteins comprise an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 2.
10. The method of claim 1 , wherein the near infrared phthalocyanine dye molecules are covalently conjugated to the papilloma virus capsid proteins.
11. The method of claim 10 , wherein the near infrared phthalocyanine dye molecules are covalently conjugated to the papilloma virus capsid proteins through covalent amide bonds.
12. The method of claim 10 , wherein the near infrared phthalocyanine dye molecules are covalently conjugated to lysine residues of the papilloma virus capsid proteins.
13. The method of claim 1 , wherein the near infrared phthalocyanine dye molecules comprise photosensitizer molecules that are promoted to an excited state upon absorption of light and undergo intersystem crossing with oxygen to produce singlet oxygen.
14. The method of claim 1 , wherein the near infrared phthalocyanine dye molecules comprise IR700 dye molecules.
15. The method of claim 1 , wherein each of the bioconjugates comprises about 50 to about 500 near infrared phthalocyanine dye molecules.
16. The method of claim 15 , wherein each of the bioconjugates comprises about 200 near infrared phthalocyanine dye molecules.
17. The method of claim 15 , wherein each of the bioconjugates comprises about 300 near infrared phthalocyanine dye molecules.
18. The method of claim 15 , wherein each of the bioconjugates comprises about 400 near infrared phthalocyanine dye molecules.
19. The method of claim 1 wherein the cells are mammalian cells.
20. The method of claim 19 , wherein the mammalian cells are 293T cells.
21. The method of claim 19 , wherein the mammalian cells are HEK293F cells.