US5284756A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Heterodimeric osteogenic factor
Assignee: Biomet Manufacturing LLC
•
Filing: 1991-06-20
•
Granted: 1994-02-08
•
Expires: 2011-06-20
Abstract
Abstract The present invention provides osteogenically active protein preparations comprising a heterodimer of P3 OF 31-34 subunit B and P3 OF 31-34 subunit D, which subunits are linked with at least one disulfide bond and methods for their preparation. The invention further provides cell lines transformed with nucleotide sequences encoding P3 OF 31-34 subunit B and P3 OF 31-34 subunit D and vectors comprising those sequences in operative association with an expression control sequence.
Inventors
- 👤 Lynn Grinna
- 👤 Georgia Theofan
- 👤 Thomas F. Parsons
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 05:12
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/51; A; A61; A61L; A61L27/00; A61L27/14; A61L27/22; A61L27/227; A61P; A61P43/00; C07K16/00; C07K16/18; C07K16/22; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; A61K; A61K38/00
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
138 words
A method of producing an osteogenic protein preparation comprising a heterodimer of a first polypeptide subunit and a second polypeptide subunit comprising the step of culturing in a suitable culture media a cell line transformed with a first and a second nucleotide sequence, said first nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 3; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 3; and said second nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 1; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 1; to produce said heterodimer.
Claim 2
Independent Claim
138 words
A method of producing an osteogenic protein preparation comprising a heterodimer of a first polypeptide subunit and a second polypeptide subunit comprising the step of culturing in a suitable culture media a cell line transformed with a first and a second nucleotide sequence, said first nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 3; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 3; and said second nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 1; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 1; to produce said heterodimer.
Claim 3
Dependent Claim
18 words
The method of claim 1 wherein said polypeptide subunits are expressed by means of a heterologous prepro sequence.
Claim 4
Dependent Claim
23 words
The method of claim 1 wherein said cell line is selected from the group consisting of mammalian, bacterial, insect and yeast cell lines.
Claim 5
Dependent Claim
15 words
The method of claim 1 wherein said cell line is Chinese hamster ovary cell lines.
Claim 6
Dependent Claim
84 words
The method of claim 1 wherein said heterodimer is isolated from the culture medium according to the steps of (a) subjecting said culture medium to a series of chromatography steps utilizing a Q-Sepharose column, an S-Sepharose column and a Phenyl-Sepharose column to recover an active fraction and (b) subjecting the active fraction to reverse phase chromatography using a C-18 high performance liquid chromatography column equilibrated with buffers containing trifluoroacetic acid and acetonitrile by eluting the active preparation at concentrations between 35% and 45% acetonitrile.
Claim 7
Dependent Claim
108 words
A cell transformed with purified and isolated nucleic acids comprising a first and a second nucleotide sequence, said first nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 3; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 3; and said second nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 1; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 1.
Claim 8
Dependent Claim
108 words
A cell transformed with purified and isolated nucleic acids comprising a first and a second nucleotide sequence, said first nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 3; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 3; and said second nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 1; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 1.
Claim 9
Independent Claim
134 words
A method of preparing a cell line capable of producing an osteogenic protein preparation comprising a heterodimer of a first polypeptide subunit and a second polypeptide subunit comprising the step of transforming a cell line with a first and a second nucleotide sequence, said first nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 3; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 3; and said second nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 1; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 1.
Claim 10
Independent Claim
134 words
A method of preparing a cell line capable of producing an osteogenic protein preparation comprising a heterodimer of a first polypeptide subunit and a second polypeptide subunit comprising the step of transforming a cell line with a first and a second nucleotide sequence, said first nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 3; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 3; and said second nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 1; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 1.
Claim 11
Dependent Claim
47 words
The method according to claim 7 comprising a first step in which said cell line is transformed with one of said first and second nucleotide sequences and a second step in which said cell line is transformed with the other of said first and second nucleotide sequences.
Claim 12
Dependent Claim
24 words
The method according to claim 7 wherein said cell line is transformed with a vector comprising both said first and said second nucleotide sequences.
Claim 13
Dependent Claim
102 words
A expression vector comprising a first and a second nucleotide sequence said first nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 3; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 3; and said second nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 1; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 1.
Claim 14
Dependent Claim
102 words
A expression vector comprising a first and a second nucleotide sequence said first nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 3; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 3; and said second nucleotide sequence being selected from the group consisting of: the nucleotide sequence as shown in SEQ ID NO: 1; and a nucleotide sequence which encodes the same sequence of amino acids as encoded by the nucleotide sequence shown in SEQ ID NO: 1.
References Cited (Backward Citations)
US3458397A
Cited
US4488911A
Cited
US4500707A
Cited
US4294753A
Cited
US4761471A
Cited
US4789732A
Cited
US4314380A
Cited
US4407787A
Cited
US4455256A
Cited
US4619989A
Cited
US4394370A
Cited
US4430760A
Cited
US4440750A
Cited
US4434094A
Cited
US4863899A
Cited
US4757006A
Cited
US4804744A
Cited
US4563489A
Cited
US4608199A
Cited
US4774228A
Cited
EP0169016A2
Cited
US4627982A
Cited
US4774322A
Cited
US4563350A
Cited
US5001169A
Cited
US4734363A
Cited
US4681763A
Cited
EP0212474A2
Cited
US4725536A
Cited
US5108922A
Cited
US5187076A
Cited
US5166058A
Cited
US5141905A
Cited
US5116738A
Cited
US5106748A
Cited
WO1988000205A1
Cited
US5013649A
Cited
US4877864A
Cited
WO1988007548A1
Cited
US4952404A
Cited
US5017486A
Cited
WO1989009787A2
Cited
WO1989009788A2
Cited
WO1989010409A1
Cited
US4968590A
Cited
EP0349048A2
Cited
US5011691A
Cited
US4975526A
Cited
WO1990011366A1
Cited
EP0409472A1
Cited
WO1991002744A1
Cited
EP0416578A2
Cited
WO1991005802A1
Cited
US5168050A
Cited
WO1992005199A1
Cited
WO1992015323A1
Cited
WO1992019262A1
Cited
WO1993000432A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1997-02-07 | 1997-08-09 | 1998-02-08 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2001-02-07 | 2001-08-09 | 2002-02-08 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2005-02-07 | 2005-08-09 | 2006-02-08 | $7700.00 | Window Closed / Lapsed |