Methods for producing secreted ligand-binding fusion proteins
DCFirst Claim
1. A method for producing a secreted ligand-binding fusion protein comprising:
- (a) introducing into a host cell a DNA construct comprising a transcriptional promoter operatively linked to a secretory signal sequence followed downstream by and in proper reading frame with a DNA sequence encoding a polypeptide chain that comprises (i) a ligand-binding domain of a receptor, wherein the receptor is a receptor for a growth factor or a hormone, and (ii) an immunoglobulin constant region polypeptide; and
(b) culturing the host cell in an appropriate culture medium whereby the ligand-binding fusion protein is secreted, wherein the ligand-binding fusion protein binds the growth factor or the hormone.
0 Assignments
Litigations
0 Petitions
Accused Products
Abstract
Methods for producing secreted receptor analogs and biologically active peptide dimers are disclosed. The methods for producing secreted receptor analogs and biologically active peptide dimers utilize a DNA sequence encoding a receptor analog or a peptide requiring dimerization for biological activity joined to a dimerizing protein. The receptor analog includes a ligand-binding domain. Polypeptides comprising essentially the extracellular domain of a human PDGF receptor fused to dimerizing proteins, the portion being capable of binding human PDGF or an isoform thereof, are also disclosed. The polypeptides may be used within methods for determining the presence of and for purifying human PDGF or isoforms thereof.
122 Citations
9 Claims
-
1. A method for producing a secreted ligand-binding fusion protein comprising:
-
(a) introducing into a host cell a DNA construct comprising a transcriptional promoter operatively linked to a secretory signal sequence followed downstream by and in proper reading frame with a DNA sequence encoding a polypeptide chain that comprises (i) a ligand-binding domain of a receptor, wherein the receptor is a receptor for a growth factor or a hormone, and (ii) an immunoglobulin constant region polypeptide; and
(b) culturing the host cell in an appropriate culture medium whereby the ligand-binding fusion protein is secreted, wherein the ligand-binding fusion protein binds the growth factor or the hormone. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
(c) obtaining the ligand-binding fusion protein from the culture medium of the cultured host cell.
-
-
3. The method of claim 1, wherein the ligand binding fusion protein competitively binds the growth factor or the hormone.
-
4. The method of claim 1, wherein the immunoglobulin constant region polypeptide comprises an immunoglobulin heavy chain constant region domain.
-
5. The method of claim 4, wherein the immunoglobulin heavy chain constant region domain is an IgG heavy chain constant region domain.
-
6. The method of claim 4, wherein the polypeptide chain further comprises an immunoglobulin hinge region joined to the immunoglobulin heavy chain constant region domain.
-
7. The method of claim 1, wherein the ligand-binding fusion protein is a dimer.
-
8. The method of claim 7, wherein the dimer is a homodimer.
-
9. A method for producing a secreted ligand-binding fusion protein comprising:
-
(a) culturing a host cell in an appropriate culture medium, wherein the host cell comprises a DNA construct comprising a transcriptional promoter operatively linked to a secretory signal sequence followed downstream by and in proper reading frame with a DNA sequence encoding a polypeptide chain that comprises (i) a ligand-binding domain of a receptor, wherein the receptor, and (ii) an immunoglobulin constant region polypeptide, whereby the ligand-binding fusion protein is secreted; and
(b) obtaining the ligand-binding fusion protein from the medium of the cultured host cell, wherein the ligand-binding fusion protein binds the growth factor or the hormone.
-
Specification