IMMUNOGLOBULIN FUSION PROTEINS AND USES THEREOF
A fusion protein is disclosed. The fusion protein of the invention comprises an Fc fragment of an immunoglobulin G and a bioactive molecule, wherein the Fc is a single chain Fc. The amino acids in the hinge of the Fc is mutated, substituted, or deleted so that the hinge of Fc cannot form disulfide bonds. Methods for producing and using the fusion protein of the invention are also provided.
1 . A method for producing a fusion protein comprising:
a) providing a bioactive molecule, an Fc fragment of an IgG molecule, and a hinge region; and
b) combing the bioactive molecule and the Fc fragment through the hinge region;
wherein a molecular weight of the bioactive molecule is about 9,000 to about 70,000 daltons, the Fc fragment comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 61-64, and the hinge region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-60.
2 . The method according to claim 1 , wherein the Fc fragment is a single chain Fc (sFc).
3 . The method according to claim 1 , wherein the hinge region is mutated and does not form a disulfide bond.
4 . The method according to claim 1 , wherein the bioactive molecule is a glycoprotein.
5 . The method according to claim 1 , wherein the bioactive molecule is a cytokine, a growth factor, a hormone, or a functional portion thereof.
6 . The method according to claim 1 , wherein the bioactive molecule is erythropoietin, Factor IX, IFNα, GCSF and IFNβ.
7 . The method according to claim 1 , wherein the hinge region comprises an amino acid sequence of SEQ ID NO: 23 or 27.
8 . The method according to claim 1 , wherein the bioactive molecule comprises an amino acid sequence of SEQ ID NO: 65.
9 . The method according to claim 1 , wherein the bioactive molecule comprises an amino acid sequence of SEQ ID NO: 67.
10 . The method according to claim 1 , wherein the bioactive molecule comprises an amino acid sequence of SEQ ID NO: 69.
11 . The method according to claim 1 , wherein the bioactive molecule comprises an amino acid sequence of SEQ ID NO: 71.
12 . The method according to claim 1 , wherein the bioactive molecule comprises an amino acid sequence of SEQ ID NO: 73.
13 . The method according to claim 1 , wherein the amino acid sequence of the fusion protein is SEQ ID NO: 66.
14 . The method according to claim 1 , wherein the amino acid sequence of the fusion protein is SEQ ID NO: 68.
15 . The method according to claim 1 , wherein the amino acid sequence of the fusion protein is SEQ ID NO: 70.
16 . The method according to claim 1 , wherein the amino acid sequence of the fusion protein is SEQ ID NO: 72.
17 . The method according to claim 1 , wherein the amino acid sequence of the fusion protein is SEQ ID NO: 74.
18 . The method according to claim 1 , wherein the bioactive molecule is directly linked to the hinge region.
19 . The method according to claim 1 , wherein step a) further comprises providing a linker, and the bioactive molecule is linked to the hinge region through the linker.
20 . The method according to claim 19 , wherein the linker comprises about 1-5 amino acids.