Recombinant bi-functional fusion protein and preparation and application therefor
The present invention provides a recombinant fusion protein containing a first extracellular Ig-like domain of a signal-regulator protein alpha (SIRPα), linked to an Fc fragment of a human IgG1. The present invention also provides a polynucleotide encoding the recombinant fusion protein, an expression vector containing the polynucleotide, a method for producing the recombinant protein and a method for treating a disease caused by over expression of CD47.
1. A recombinant bi-functional fusion protein, comprising an extracellular Ig-like domain of a signal-regulator protein (SIRP), linked to an Fc region of an immunoglobulin comprising both CH2 and CH3 regions of the immunoglobulin, wherein the signal-regulatory protein is SIRPα, the extracellular Ig-like domain of the signal-regulator protein is, and the SIRPαD1 has mutation N89A and consists of the amino acid sequence from residue 1 to 142 of SEQ ID NO.: 6, wherein the recombinant bi-functional fusion protein can bind to Cluster of Differentiation 47 (CD47) and Fcγ Receptor, blocking the binding of CD47 to the SIRP on the cell surface of macrophages and stimulating phagocytosis of tumor cells by macrophages.
2. The recombinant bi-functional fusion protein of claim 1 , wherein the Fc region is an Fc region of IgG1.
3. The recombinant bi-functional fusion protein of claim 2 , wherein the IgG1 is a human IgG1.
4. A homodimer comprising two recombinant bi-functional fusion proteins of claim 1 linked by one or more disulfide bonds.
5. The recombinant bi-functional fusion protein of claim 3 , wherein the Fc region comprises an amino acid sequence having at least 95% identity to the amino acid sequence set forth in SEQ ID NO: 6 from residue 143 to residue 374.
6. The recombinant bi-functional fusion protein of claim 3 , wherein the Fc region comprises an amino acid sequence having at least 98% identity to the amino acid sequence set forth in SEQ ID NO: 6 from residue 143 to residue 374.
7. The recombinant bi-functional fusion protein of claim 3 , wherein the Fc region comprises an amino acid sequence having at least 99% identity to the amino acid sequence set forth in SEQ ID NO: 6 from residue 143 to residue 374.
8. The recombinant bi-functional fusion protein of claim 5 , wherein the Fc region consists of the amino acid sequence from residue 143 to residue 374 of SEQ ID NO.: 6.
9. The recombinant bi-functional fusion protein of claim 8 , comprising the amino acid sequence set forth in SEQ ID NO.: 6.
10. The recombinant bi-functional fusion protein of claim 9 , consisting of the amino acid sequence set forth in SEQ ID NO.: 6.
11. The homodimer of claim 4 , wherein the Fc region is an Fc region of IgG1.
12. The homodimer of claim 11 , wherein the IgG1 is a human IgG1.
13. The homodimer of claim 12 , wherein the Fc region consists of the amino acid sequence from residue 143 to residue 374 of SEQ ID NO.: 6.