Synthetic membrane-receiver complexes
Compositions comprising synthetic membrane-receiver complexes, methods of generating synthetic membrane-receiver complexes, and methods of treating or preventing diseases, disorders or conditions therewith.
1. An enucleated erythroid cell comprising on its surface at least 1,000 copies of an exogenous polypeptide comprising asparaginase or a functional fragment thereof, wherein the enucleated erythroid cell was made by a process comprising introducing an exogenous nucleic acid encoding the exogenous polypeptide into a nucleated erythroid cell precursor.
2. The enucleated erythroid cell of claim 1 , which comprises on its surface at least 10,000 copies of the exogenous polypeptide.
3. The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide further comprises a transmembrane domain.
4. The enucleated erythroid cell of claim 3 , wherein the transmembrane domain comprises the transmembrane region of a type 2 membrane protein.
5. The enucleated erythroid cell of claim 3 , wherein the transmembrane domain comprises a transmembrane region of Kell.
6. The enucleated erythroid cell of claim 3 , wherein the exogenous polypeptide comprises the 71-amino acid N-terminal fragment of Kell.
7. The enucleated erythroid cell of claim 3 , wherein the exogenous polypeptide comprises the 79-amino acid N-terminal fragment of Kell.
8. The enucleated erythroid cell of claim 3 , wherein the asparaginase or functional fragment thereof is fused to the C-terminus of the transmembrane domain.
9. The enucleated erythroid cell of claim 8 , wherein a linker is disposed between the C terminus of the transmembrane domain and the asparaginase or functional fragment thereof.
10. The enucleated erythroid cell of claim 9 , wherein the linker comprises a poly-glycine poly-serine linker.
11. The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide comprises either Escherichia coli asparaginase or a functional fragment thereof or Erwinia chrysanthemi asparaginase or a functional fragment thereof.
12. The enucleated erythroid cell of claim 1 , which is a reticulocyte.
13. The enucleated erythroid cell of claim 1 , wherein the enucleated erythroid cell exhibits substantially the same osmotic membrane fragility as an isolated, unmodified, cultured or uncultured erythroid cell.
14. The enucleated erythroid cell of claim 1 , which further comprises a second exogenous polypeptide on the cell surface.
15. The enucleated erythroid cell of claim 14 , wherein the second exogenous polypeptide is a fusion polypeptide.
16. The enucleated erythroid cell of claim 14 , wherein the second exogenous polypeptide comprises a transmembrane domain.
17. The enucleated erythroid cell of claim 14 , wherein the second exogenous polypeptide comprises an antibody.
18. The enucleated erythroid cell of claim 17 , wherein the antibody is a single chain antibody.
19. The enucleated erythroid cell of claim 14 , wherein the second exogenous polypeptide binds a cancer cell.
20. A nucleated erythroid cell precursor comprising at least 1,000 copies of a surface exposed exogenous polypeptide comprising asparaginase or a functional fragment thereof, wherein the nucleated erythroid cell precursor was made by a process comprising introducing an exogenous nucleic acid encoding the exogenous polypeptide into the nucleated erythroid cell precursor.
21. The enucleated erythroid cell of claim 1 , which is a mature erythrocyte.
22. The enucleated erythroid cell of claim 1 , wherein the exogenous nucleic acid comprises DNA.
23. The enucleated erythroid cell of claim 1 , wherein the exogenous nucleic acid comprises RNA.
24. The enucleated erythroid cell of claim 1 , which lacks A and B antigen.
25. The enucleated erythroid cell of claim 1 , which comprises fetal hemoglobin.
26. The enucleated erythroid cell of claim 1 , which is not hypotonically loaded.
27. A pharmaceutical composition comprising a plurality of enucleated erythroid cells, wherein each of the enucleated erythroid cell of said plurality comprises on its surface at least 1,000 copies of an exogenous polypeptide comprising asparaginase or a functional fragment thereof, wherein each of the enucleated erythroid cells of said plurality was made by a process comprising introducing into an erythroid cell precursor an exogenous nucleic acid encoding the exogenous polypeptide.
28. The pharmaceutical composition of claim 27 , wherein at least 50% of enucleated erythroid cells in the pharmaceutical composition comprise the exogenous polypeptide.
29. The pharmaceutical composition of claim 27 , wherein at least 60% of cells in the pharmaceutical composition are enucleated erythroid cells.