US6486303B1
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Method for making hormone heterodimers
Assignee: NEW JERSEY UNIVERSITY OF MEDICINE & DENTISTRY OF
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Filing: 1998-04-14
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Granted: 2002-11-26
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Expires: 2018-04-14
Abstract
Abstract The present invention relates to a method for preparing heterodimeric analogs of cysteine knot proteins. More specifically, the invention relates to a method for forming a subunit combination of a cysteine knot protein having an alpha-subunit and a beta-subunit to prepare a heterodimeric protein analog which comprises the steps of (a) attaching a dimerization domain to the amino termini of both an alpha-subunit and beta-subunit of a cysteine knot protein; and (b) dimerizing the alpha-subunit and beta-subunit to form a heterodimeric protein analog. In another embodiment, the invention relates to a method for forming a subunit combination of a cysteine knot protein having an alpha-subunit and a beta-subunit to prepare a heterodimeric protein analog which comprises the steps of (a) attaching a dimerization domain to the amino terminus of an alpha-subunit and the carboxy terminus of a beta-subunit of a cysteine knot protein; and (b) dimerizing the alpha-subunit and beta-subunit to form a heterodimeric protein analog.
Inventors
- 👤 William R. Moyle
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 16:32
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/59; C07K14/82; C07K2319/00
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
65 words
A method for forming a subunit combination of a cysteine knot glycoprotein hormone having an α-subunit and a β-subunit to prepare a heterodimeric protein analog which comprises the steps of: (a) attaching a dimerization domain to the amino termini of both an α-subunit and a β-subunit of a cysteine knot glycoprotein hormone; and (b) dimerizing the α-subunit and β-subunit to form a heterodimeric protein analog.
Claim 2
Independent Claim
65 words
A method for forming a subunit combination of a cysteine knot glycoprotein hormone having an α-subunit and a β-subunit to prepare a heterodimeric protein analog which comprises the steps of: (a) attaching a dimerization domain to the amino termini of both an α-subunit and a β-subunit of a cysteine knot glycoprotein hormone; and (b) dimerizing the α-subunit and β-subunit to form a heterodimeric protein analog.
Claim 3
Dependent Claim
35 words
The method according to claim 1 , wherein a Fos dimerization sequence domain is attached to the amino-terminus of the β-subunit and a Jun dimerization sequence domain is attached to the amino-terminus of the α-subunit.
Claim 4
Dependent Claim
35 words
The method according to claim 1 , wherein a Fos dimerization sequence domain is attached to the amino-terminus of the α-subunit and a Jun dimerization sequence domain is attached to the amino-terminus of the β-subunit.
Claim 5
Dependent Claim
39 words
The method according to claim 2 , wherein a glycine or serine residue is inserted between the Fos or Jun dimerization sequence domain and a furin cleavage site to facilitate cleavage of the dimerization sequence domain from the heterodimer.
Claim 6
Dependent Claim
39 words
The method according to claim 3 , wherein a glycine or serine residue is inserted between the Fos or Jun dimerization sequence domain and a furin cleavage site to facilitate cleavage of the dimerization sequence domain from the heterodimer.
Claim 7
Dependent Claim
39 words
The method according to claim 1 , wherein the heterodimeric protein analog is selected from the group consisting of hCG/hFSH chimeras, hCG/hTSH chimeras, deglycosylated hormones, truncated glycoprotein hormones, mutant glycoprotein hormones, and glycoprotein hormones containing an hCG carboxyl terminus.
Claim 8
Dependent Claim
38 words
The method according to claim 1 , further comprising incorporating protease cleavage sites between additional N-terminal sequences and the α-subunit and the β-subunit of the cysteine knot protein to remove the dimerization domains from the heterodimeric protein analog.
Claim 9
Dependent Claim
23 words
The method according to claim 1 , wherein the cysteine knot glycoprotein hormone has an oligosaccharide genetically removed from the α-subunit at Asn52.
Claim 10
Dependent Claim
16 words
The method according to claim 1 , wherein the cysteine knot glycoprotein hormone lacks a seatbelt.
Claim 11
Independent Claim
68 words
A method for forming a subunit combination of a cysteine knot glycoprotein hormone having an α-subunit and a β-subunit to prepare a heterodimeric protein analog which comprises the steps of: (a) attaching a dimerization domain to the amino terminus of an α-subunit and the carboxy terminus of a β-subunit of a cysteine knot glycoprotein hormone; and (b) dimerizing the α-subunit and β-subunit to form a heterodimeric protein analog.
Claim 12
Independent Claim
68 words
A method for forming a subunit combination of a cysteine knot glycoprotein hormone having an α-subunit and a β-subunit to prepare a heterodimeric protein analog which comprises the steps of: (a) attaching a dimerization domain to the amino terminus of an α-subunit and the carboxy terminus of a β-subunit of a cysteine knot glycoprotein hormone; and (b) dimerizing the α-subunit and β-subunit to form a heterodimeric protein analog.
Claim 13
Dependent Claim
35 words
The method according to claim 10 , wherein a Fos dimerization sequence domain is attached to the carboxy-terminus of the β-subunit and a Jun dimerization sequence domain is attached to the amino-terminus of the α-subunit.
Claim 14
Dependent Claim
36 words
The method according to claim 10 , wherein a Fos dimerization sequence domain is attached to the amino-terminus of the α-subunit and a Jun dimerization sequence domain is attached to the carboxy terminus of the β-subunit.
Claim 15
Dependent Claim
37 words
The method according to claim 11 , wherein a protease cleavage site is inserted between the dimerization sequence domain and the α-subunit and a protease cleavage site is inserted between the dimerization sequence domain and the β-subunit.
Claim 16
Dependent Claim
14 words
The method according to claim 13 , wherein the protease cleavage site is furin.
Claim 17
Dependent Claim
39 words
The method according to claim 10 , wherein the heterodimeric protein analog is selected from the group consisting of hCG/hFSH chimeras, hCG/hTSH chimeras, deglycosylated hormones, truncated glycoprotein hormones, mutant glycoprotein hormones, and glycoprotein hormones containing an hCG carboxyl terminus.
Claim 18
Dependent Claim
23 words
The method according to claim 10 , wherein the cysteine knot glycoprotein hormone has an oligosaccharide genetically removed from the α-subunit at Asn52.
Claim 19
Dependent Claim
37 words
The method according to claim 1 , wherein a Fos dimerization sequence domain is attached to the amino-terminus of an hCG β-subunit and a Jun dimerization sequence domain is attached to the amino-terminus of an hFSH α-subunit.
Claim 20
Dependent Claim
37 words
The method according to claim 10 , wherein a Fos dimerization sequence domain is attached to the carboxy-terminus of an hCG β-subunit and a Jun dimerization sequence domain is attached to the amino-terminus of an hFSH α-subunit.
References Cited (Backward Citations)
WO1991016922A1
Cited
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