Aptamer modulators of complement protein C3 and biologically active proteolytic products thereof
This invention relates to nucleic acid aptamers that recognize and bind the complement protein C3 or its biologically active proteolytic products and methods of their use. Particularly preferred are bi-functional aptamer construct that binding specifically with C3b or iC3b, and another target protein. Use of these molecular constructs for commandeering the opsonization process is also described herein.
1. A nucleic acid aptamer comprising:
a first domain that binds to a biologically active proteolytic product of complement protein C3 that is C3b, iC3b, or both thereof, wherein the first domain binds to a site on C3b or iC3b that is distinct of, respectively, a CR1 or CR3 receptor binding site on C3b or iC3b, wherein the first domain comprises a nucleotide sequence according to nucleotides 2-41 of SEQ ID NO: 9; and
a second domain that binds to a molecule different from the biologically active proteolytic product of complement protein C3.
2. The nucleic acid aptamer according to claim 1 further comprising:
a plurality of first domains that bind to the biologically active proteolytic product of complement protein C3; or
a plurality of second domains that bind to a molecule different from the biologically active proteolytic product of complement protein C3.
3. The nucleic acid aptamer according to claim 1 , wherein the biologically active proteolytic product of complement protein C3 is one of C3b and iC3b.
4. The nucleic acid aptamer according to claim 1 , wherein the molecule different from the biologically active proteolytic product of complement protein C3 is a target protein.
5. The nucleic acid aptamer according to claim 4 , wherein the target protein is a cell surface protein.
6. The nucleic acid aptamer according to claim 4 , wherein the target protein is an extracellular protein.
7. The nucleic acid aptamer according to claim 1 further comprising a near-infrared emitting probe conjugated to the aptamer.
8. A molecular complex comprising:
the nucleic acid aptamer according to claim 1 ; and
the biologically active proteolytic product of complement protein C3 bound to the nucleic acid aptamer at the first domain.
9. The molecular complex according to claim 8 , wherein the biologically active proteolytic product of complement protein C3 is one of C3b and iC3b.
10. The molecular complex according to claim 8 further comprising:
a target protein bound to the nucleic acid aptamer at the second domain.
11. The molecular complex according to claim 10 , wherein the target protein is a cell surface protein.
12. The molecular complex according to claim 11 , wherein the cell surface protein is a cancer cell-specific surface protein.
13. The molecular complex according to claim 10 , wherein the target protein is an extracellular protein.
14. The molecular complex according to claim 13 , wherein the extracellular protein is VEGF.
15. The molecular complex according to claim 10 , wherein the target protein is a fluorescent protein.
16. A cell comprising a target protein on its surface, wherein the target protein is a component of the molecular complex according to claim 10 .
17. The cell according to claim 16 , wherein the cell is a cancer cell.
18. A method of promoting opsonization of a cell, said method comprising:
providing a nucleic acid aptamer according to claim 1 ; and
contacting a cell surface protein on a cell with the aptamer, wherein the aptamer binds the target protein at the second domain and the biologically active proteolytic product of complement protein C3 at the first domain, thereby promoting opsonization of the cell.
19. The method according to claim 18 , wherein the cell is a cancer cell and the cell surface protein is cancer cell-specific.
20. The nucleic acid aptamer according to claim 1 wherein the nucleic acid aptamer comprises 2′-fluoro RNA.