DESC:CHIMERIC PLANT BASED VIRUS-LIKE-PARTICLES, AND USES THEREOF ,CLAIMS:1. A virus-like-particle (VLP) comprising:
a. a first viral coat protein (CP) having amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 117 and SEQ ID NO: 119 and,
b. at least one Staphylococcus protein A (SpA) domain having amino acid sequence selected from the group consisting of SEQ ID NO: 7, and SEQ ID NO: 9,
wherein said first viral CP is fused to said at least one SpA domain.
2. The VLP as claimed in claim 1, wherein
a. said at least one SpA domain is fused to N-terminal end of said first viral CP; or
b. said at least one SpA domain is fused to C-terminal end of said first viral CP; or
c. said at least one SpA domain is integrated within the said first viral CP; or
d. said at least one SpA domain is integrated at N and C termini of said first viral CP; or
e. said at least one SpA domain is integrated at N-terminal, C-terminal and within said first viral CP; or
f. said at least one SpA domain is integrated at N-terminal and within said first viral CP; or
g. said at least one SpA domain is integrated at C-terminal and within said first viral CP.
3. The VLP as claimed in any of the claims 1-2, wherein said first viral CP is encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 118 and SEQ ID NO: 120 and said at least one SpA domain is encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NO: 8, and SEQ ID NO: 10.
4. A virus-like-particle (VLP) having amino acid sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO:SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO:113, SEQ ID NO:115, SEQ ID NO:121, SEQ ID NO:123, SEQ ID NO:125, SEQ ID NO:127, SEQ ID NO:129, SEQ ID NO:131, SEQ ID NO:133, SEQ ID NO:135, SEQ ID NO:137, SEQ ID NO:139, SEQ ID NO:141, SEQ ID NO:143, SEQ ID NO:145, SEQ ID NO:147, SEQ ID NO:149, SEQ ID NO:151, SEQ ID NO:153, SEQ ID NO:155, SEQ ID NO:157, and SEQ ID NO:159.
5. The VLP as claimed in claim 4, wherein said VLP is encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112. SEQ ID NO:114, SEQ ID NO:116, SEQ ID NO:122, SEQ ID NO:124, SEQ ID NO:126, SEQ ID NO:128, SEQ ID NO:130, SEQ ID NO:132, SEQ ID NO:134, SEQ ID NO:136, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, SEQ ID NO:144, SEQ ID NO:146, SEQ ID NO:148, SEQ ID NO:150, SEQ ID NO:152, SEQ ID NO:154, SEQ ID NO:156, SEQ ID NO:158 and SEQ ID NO:160.
6. A DNA construct comprising a polynucleotide fragment operably linked to a promoter, wherein said polynucleotide fragment encodes a virus-like-particle (VLP) comprising:
a. a first viral coat protein (CP) having amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 117 and SEQ ID NO: 119 and
b. at least one Staphylococcus protein A (SpA) domain having amino acid sequence selected from the group consisting of SEQ ID NO: 7, and SEQ ID NO: 9,
wherein said first viral CP is fused to said at least one SpA domain.
7. The DNA construct as claimed in claim 6, wherein said VLP amino acid sequence is selected from the group consisting of SEQ ID NO: 11, SEQ ID NO:SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO:113, SEQ ID NO:115, SEQ ID NO:121, SEQ ID NO:123, SEQ ID NO:125, SEQ ID NO:127, SEQ ID NO:129, SEQ ID NO:131, SEQ ID NO:133, SEQ ID NO:135, SEQ ID NO:137, SEQ ID NO:139, SEQ ID NO:141, SEQ ID NO:143, SEQ ID NO:145, SEQ ID NO:147, SEQ ID NO:149, SEQ ID NO:151, SEQ ID NO:153, SEQ ID NO:155, SEQ ID NO:157, and SEQ ID NO:159.
8. The DNA construct as claimed in claim 6, wherein said polynucleotide fragment sequence is selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, and SEQ ID NO: 112, SEQ ID NO:114, SEQ ID NO:116, SEQ ID NO:122, SEQ ID NO:124, SEQ ID NO:126, SEQ ID NO:128, SEQ ID NO:130, SEQ ID NO:132, SEQ ID NO:134, SEQ ID NO:136, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, SEQ ID NO:144, SEQ ID NO:146, SEQ ID NO:148, SEQ ID NO:150, SEQ ID NO:152, SEQ ID NO:154, SEQ ID NO:156, SEQ ID NO:158 and SEQ ID NO:160.
9. A recombinant DNA vector comprising a DNA construct, said DNA construct as claimed in any of the claims 6-8.
10. A recombinant host cell comprising a DNA construct as claimed in any of the claims 6-8.
11. A recombinant host cell comprising a recombinant DNA vector as claimed in claim 9.
12. The recombinant host cell as claimed in any of the claim 10-11, wherein said recombinant host cell is of bacterial, fungal, plant, insect, or mammalian origin.
13. A process for purifying an immunoglobulin comprising the steps:
a. obtaining a virus-like-particle (VLP) as claimed in any of the claims 1-5;
b. contacting said VLP with an immunoglobulin under conditions which allow binding of said VLP with said immunoglobulin; and
c. separating said immunoglobulin from said VLP.
14. A process for delivery of antibodies to the surface, inside or both to surface and inside of cells, said process comprising the steps:
a. obtaining a virus-like-particle (VLP) as claimed in any of the claims 1-5;
b. obtaining at least one antibody;
c. incubating said VLP with said at least one antibody to obtain VLP-antibody complexes; and
d. contacting said VLP-antibody complexes with said cells.
15. A process for imaging a tissue of interest, said process comprising the steps:
a. obtaining a virus-like-particle (VLP) as claimed in any of the claims 1-5;
b. obtaining at least one antibody;
c. labelling said VLP and said at least one antibody with a fluorophore, wherein the fluorophore for labelling said VLP is different than the fluorophore for labelling said at least one antibody;
d. Incubating said labelled VLP with said labelled at least one antibody to obtain VLP-antibody complexes;
e. administering said VLP-antibody complex to a subject; and
f. visualizing said tissue of interest by any known method.
16. A process of targeting a virus-like-particle (VLP) to a cancer cell, said process comprising the steps:
a. obtaining a VLP as claimed in any of the claims 1-5;
b. obtaining at least one molecule which can ameliorate cancer cell activity;
c. obtaining at least one antibody which binds preferentially to at least one cell surface moiety present majorly in said cancer cell;
d. encapsulating at least one molecule with said VLP;
e. contacting said VLP from d) with said at least one antibody; and
f. contacting said VLP from e) with a tissue comprising cancer cell,
wherein said method targets VLP to a cancer cell.
17. The process as claimed in claim 16, further comprising conjugating said VLP from step a) or d) with at least one cell penetrating peptide.
18. A VLP as claimed in any of the claims 1-5 for use in disease diagnosis, cargo delivery, intracellular imaging, or cell specific targeting.
Dated this September 2015