Nucleic acid encoding human IL-2 variant
This invention provides for a fusion protein between an IL2αβγ Selective Agonist protein (IL2 Selective Agonist) and a IgG Fc protein using a linker. The IL2 Selective Agonist moiety provides a therapeutic activity by selectively activating the IL2αβγ form of the receptor, thus selectively stimulating Tregs. The Fc moiety provides a prolonged circulating half-life compared to the circulating half-life of IL-2 or an IL2SA protein.
1. A nucleic acid encoding a fusion protein comprising:
a. a human IL-2 variant protein domain comprising a substitution selected from the group consisting of: N88R, N88G, D20H, Q126L, and Q126F;
b. a peptide linker domain of from 5 to 30 amino acid residues, wherein the peptide linker domain comprises glycine residues, serine residues, or a mixture of glycine and serine residues; and
c. an IgG Fc protein domain,
wherein each domain has an amino-terminus (N-terminus) and a carboxy-terminus (C-terminus); and wherein the fusion protein is configured so that the C-terminus of the human IL-2 variant protein domain is fused through a peptide bond to the N-terminus of the peptide linker domain, and the N-terminus of the IgG Fc protein domain is fused through a peptide bond to the C-terminus of the peptide linker domain, wherein protein yield of the fusion protein is increased by at least two-fold relative to a fusion protein that comprises the human IL-2 variant protein domain and the IgG Fc protein domain, but does not comprise the peptide linker domain.
2. The nucleic acid of claim 1 , wherein the human IL-2 variant protein domain comprises the N88R substitution.
3. The nucleic acid of claim 1 , wherein the peptide linker domain consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18 and SEQ ID NO: 19.
4. A plasmid comprising the nucleic acid of claim 1 .
5. A host cell comprising the nucleic acid of claim 1 .
6. The host cell of claim 5 , wherein the host cell is a mammalian cell.
7. The nucleic acid of claim 1 , wherein the human IL-2 variant protein domain further comprises a C125S substitution.
8. The nucleic acid of claim 1 , wherein the peptide linker domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18 and SEQ ID NO: 19.
9. The nucleic acid of claim 1 , wherein the human IL-2 variant protein domain selectively activates the IL2Rαβγ receptor complex relative to the IL2Rβγ receptor complex.
10. A nucleic acid encoding a fusion protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 20, SEQ ID NO: 21 and SEQ ID NO: 31.
11. A plasmid comprising the nucleic acid of claim 10 .
12. A host cell comprising the nucleic acid of claim 10 .
13. The host cell of claim 12 , wherein the host cell is a mammalian cell.