IL-7 fusion proteins
View Patent ↗The invention is directed to a fusion protein which includes a first portion including an immunoglobulin (Ig) chain and a second portion including interleukin-7 (IL-7).
1. A fusion protein comprising:
a first portion comprising an Fc region from IgG1 or IgG2 wherein the Fc region comprises a hinge region, a CH2 domain, and a CH3 domain; and
a second portion comprising a polypeptide having an amino acid sequence comprising at least 95% sequence identity with wild-type mature human interleukin-7 (IL-7) wherein wild-type mature human IL-7 consists of amino acid residues 26-176 of SEQ ID NO:1, and wherein amino acid residues corresponding to positions 70 and 91 of wild-type mature human IL-7 are glycosylated and the amino acid residue corresponding to position 116 of wild-type mature human IL-7 is non-glycosylated.
2. The fusion protein of claim 1 , wherein the amino acid residue corresponding to position 116 of wild-type mature human IL-7 is asparagine.
3. The fusion protein of claim 1 , wherein the polypeptide having 95% sequence identity with wild-type mature human IL-7 comprises disulfide bonding between residues corresponding to positions Cys2 and Cys92, Cys34 and Cys 129, and Cys47 and Cys 141 of wild-type mature human IL-7.
4. The fusion protein of claim 1 , further comprising a linker between the first portion and the second portion.
5. The fusion protein of claim 1 , wherein the Fc region is from IgG1.
6. The fusion protein of claim 5 , wherein the Fc region further comprises an amino acid substituted in place of Asn297at glycosylation site of the Fc region.
7. The fusion protein of claim 6 , wherein the Fc region further comprising an amino acid substituted in place of Tyr296 at the glycosylation site of the Fc region.
8. The fusion protein of claim 7 , wherein Ala is substituted in place of Tyr296 and Gln is substituted in place of Asn297 at the glycosylation site of the Fc region.
9. The fusion protein of claim 1 , wherein the Fc region is from IgG2.
10. The fusion protein of claim 9 , wherein the Fc region further comprises an IgG1 hinge.
11. The fusion protein of claim 9 , wherein the Fc region further comprises an amino acid substituted in place of Asn at glycosylation site of the Fc region.
12. The fusion protein of claim 11 , wherein the Fc region further comprises an amino acid substituted in place of the Phe residue adjacent the Asn residue at the glycosylation site of the Fc region.
13. The fusion protein of claim 12 , wherein Ala is substituted in place of Phe and Gln is substituted in place of Asn at the glycosylation site of the Fc region.
14. The fusion protein of claim 1 , wherein said second portion comprises wild-type mature human IL-7 consisting of amino acid residues 26-176 of SEQ ID NO:1, and wherein the amino acid residues corresponding to positions 70 and 91 of said wild-type mature human IL-7 are glycosylated and the amino acid residue corresponding to position 116 of said wild-type mature human IL-7 is non-glycosylated.
15. A purified nucleic acid encoding the fusion protein of claim 1 .
16. A cultured host cell comprising the nucleic acid of claim 15 .
17. A fusion protein comprising:
a first portion comprising an Fc region from IgG1 or IgG2 wherein the Fc region comprises a hinge region, a CH2 domain, and a CH3 domain;
and a second portion comprising an amino acid sequence that is at least 95% identical to amino acid residues 26-176 of SEQ ID NO:1 (wild-type mature human IL-7) except that residues 121 to 138 of SEQ ID NO:1 are deleted in said second portion, and wherein the amino acid residues corresponding to positions 70 and 91 of wild-type mature human IL-7 are glycosylated and the amino acid residue corresponding to position 116 of wild-type mature human IL-7 is non-glycosylated.