Patent EP0376785A2 successfully refreshed from Google Patents.
EP0376785A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Cloning and expression of transforming growth factor beta 2
Assignee: Oncogen LP
•
Filing: 1989-12-14
•
Granted: —
•
Expires: 2009-12-14
Abstract
Abstract cDNA clones coding for TGF-S2 which are used to construct expression vectors capable of directing the high-level expression of mature, biologically active TGF-β2, as well as precursor TGF-β2 forms, in transfected Chinese Hamster Ovary cells (CHO cells) and transfeced COS cells are described. CHO and COS transfectants secreting TGF-β2 at high levels are also described. CHO cells transfected with a plasmid vector carrying the complete 414 amino acid simian TGF-β2 precursor secrete approximately 5 µg per ml culture media.
Inventors
- 👤 Anthony F. Purchio
- 👤 Linda Madisen
- 👤 Nancy Webb
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-24 21:59
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/495; A; A61; A61K; A61K38/00; C07K2319/00
Claims Analysis
Total Claims: 61
Claim 1
Independent Claim
32 words
A nucleotide sequence encoding transforming growth factor-β2 precursor comprising the nucleotide coding sequence substantially as depicted in FIG. 1 a from about nucleotide residue number 1 to about nucleotide residue number 1326.
Claim 2
Dependent Claim
32 words
A nucleotide sequence encoding transforming growth factor-β2 precursor comprising the nucleotide coding sequence substantially as depicted in FIG. 1 a from about nucleotide residue number 1 to about nucleotide residue number 1326.
Claim 3
Dependent Claim
53 words
A nucleotide sequence encoding transforming growth factor-β2 precursor comprising the nucleotide coding sequence substantially as depicted in FIG. 1 a from about nucleotide residue number 1 to about nucleotide residue number 1326 in which the nucleotide sequence from nucleotide 'residue numbers 346 to 432 is deleted and replaced by the nucleotide sequence "AAT".
Claim 4
Dependent Claim
53 words
A nucleotide sequence encoding transforming growth factor-β2 precursor comprising the nucleotide coding sequence substantially as depicted in FIG. 1 a from about nucleotide residue number 1 to about nucleotide residue number 1326 in which the nucleotide sequence from nucleotide 'residue numbers 346 to 432 is deleted and replaced by the nucleotide sequence "AAT".
Claim 5
Dependent Claim
30 words
A nucleotide sequence encoding mature transforming growth factor-β2 comprising the nucleotide sequence substantially as depicted in FIG. 1a from about nucleotide residue number 991 to about nucleotide residue number 1326.
Claim 6
Dependent Claim
30 words
A nucleotide sequence encoding mature transforming growth factor-β2 comprising the nucleotide sequence substantially as depicted in FIG. 1a from about nucleotide residue number 991 to about nucleotide residue number 1326.
Claim 7
Dependent Claim
30 words
A transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1a from about amino acid residue number 1 to about amino acid residue number 442.
Claim 8
Dependent Claim
30 words
A transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1a from about amino acid residue number 1 to about amino acid residue number 442.
Claim 9
Dependent Claim
51 words
A transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1a from about amino acid residue number 1 to about amino acid residue number 442 in which the amino acid sequence from residue numbers 116 to 144 is deleted and replaced by a single Asparagine residue.
Claim 10
Dependent Claim
51 words
A transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1a from about amino acid residue number 1 to about amino acid residue number 442 in which the amino acid sequence from residue numbers 116 to 144 is deleted and replaced by a single Asparagine residue.
Claim 11
Dependent Claim
30 words
Transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1a from about amino acid residue number 20 to about amino acid forming residue number 442.
Claim 12
Dependent Claim
30 words
Transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1a from about amino acid residue number 20 to about amino acid forming residue number 442.
Claim 13
Dependent Claim
49 words
Transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1a from about amino acid residue number 20 to about amino acid number 442 in which the amino acid sequence from residue numbers 116 to 144 is deleted and replaced by a single Asparagine residue.
Claim 14
Dependent Claim
49 words
Transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1a from about amino acid residue number 20 to about amino acid number 442 in which the amino acid sequence from residue numbers 116 to 144 is deleted and replaced by a single Asparagine residue.
Claim 15
Dependent Claim
29 words
Transforming growth factor-β2 comprising the amino acid sequence substantially as depicted in FIG. 1 a from about amino acid residue number 331 to about amino acid residue number 442.
Claim 16
Dependent Claim
29 words
Transforming growth factor-β2 comprising the amino acid sequence substantially as depicted in FIG. 1 a from about amino acid residue number 331 to about amino acid residue number 442.
Claim 17
Dependent Claim
31 words
A nucleotide sequence encoding transforming growth factor-02 precursor comprising the nucleotide coding sequence substantially as depicted in FIG. 1 from about nucleotide residue number -70 to about nucleotide residue number 1755.
Claim 18
Dependent Claim
31 words
A nucleotide sequence encoding transforming growth factor-02 precursor comprising the nucleotide coding sequence substantially as depicted in FIG. 1 from about nucleotide residue number -70 to about nucleotide residue number 1755.
Claim 19
Dependent Claim
31 words
A transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1 b from about amino acid residue number 1 to about amino acid residue number 390.
Claim 20
Dependent Claim
31 words
A transforming growth factor-β2 precursor comprising the amino acid sequence substantially as depicted in FIG. 1 b from about amino acid residue number 1 to about amino acid residue number 390.
Claim 21
Dependent Claim
31 words
A transforming growth factor-02 precursor comprising the amino acid sequence substantially as depicted in FIG. 1 b from about amino acid residue number 30 to about amino acid residue number 390.
Claim 22
Dependent Claim
31 words
A transforming growth factor-02 precursor comprising the amino acid sequence substantially as depicted in FIG. 1 b from about amino acid residue number 30 to about amino acid residue number 390.
Claim 23
Dependent Claim
30 words
Transforming growth factor β2 comprising the amino acid sequence substantially as depicted in FIG. 1 b from about amino acid residue number 279 to about amino acid residue number 390.
Claim 24
Dependent Claim
30 words
Transforming growth factor β2 comprising the amino acid sequence substantially as depicted in FIG. 1 b from about amino acid residue number 279 to about amino acid residue number 390.
Claim 25
Independent Claim
60 words
A method for producing transforming growth factor-β2 comprising: (a) culturing a eucaryotic cell containing a nucleotide sequence encoding transforming growth factor-β2 under the control of a second nucleotide sequence that regulates gene expression so that a peptide or protein having transforming growth factor-02 activity is produced by the eucaryotic cell; and (b) recovering the transforming growth factor-β2 from the culture.
Claim 26
Independent Claim
60 words
A method for producing transforming growth factor-β2 comprising: (a) culturing a eucaryotic cell containing a nucleotide sequence encoding transforming growth factor-β2 under the control of a second nucleotide sequence that regulates gene expression so that a peptide or protein having transforming growth factor-02 activity is produced by the eucaryotic cell; and (b) recovering the transforming growth factor-β2 from the culture.
Claim 27
Dependent Claim
31 words
The method according to claim 13 in which the nucleotide sequence encoding transforming growth factor-82 comprises the nucleotide sequence substantially as depicted in FIG. 1a from nucleotide number 1 to 1339.
Claim 28
Dependent Claim
32 words
.15. The method according to claim 13 in which the nucleotide sequence encoding transforming growth factor-β2 comprises the nucleotide sequence substantially as depicted in FIG. 1 from nucleotide number -70 to 1755.
Claim 29
Dependent Claim
17 words
The method according to claim 13 in which the eucaryotic cell comprises a Chinese Hamster Ovary cell.
Claim 30
Dependent Claim
15 words
The method according to claim 13 in which the eucaryotic cell comprises a COS cell.
Claim 31
Dependent Claim
20 words
The method according to claim 13 in which the second nucleotide sequence which controls gene expression comprises an SV40 promoter.
Claim 32
Dependent Claim
44 words
The method according to claim 13 in which the second nucleotide sequence comprises a promoter and a sequence encoding a selectable marker for which the eucaryotic cell is deficient, so that the eucaryotic cell containing the transforming growth factor-β2 coding sequence can be identified.
Claim 33
Dependent Claim
14 words
The method according to claim 19 in which the selectable marker comprises dihydrofolate reductase.
Claim 34
Dependent Claim
35 words
The method according to claim 20 further comprising exposing the eucaryotic cell to methotrexate, so that resistant colonies are selected which contain amplified levels of the coding sequences for dihydrofolate reductase and transforming growth factor-,82.
Claim 35
Dependent Claim
19 words
The method according to claim 21 in which the eucaryotic cell comprises a dihydrofolate reductase-deficient Chinese Hamster Ovary cell.
Claim 36
Independent Claim
36 words
A method for producing transforming growth factor-β2, comprising (a) culturing transfectant CHO-1/39, 12.5, CL 36 as deposited with the ATCC and assigned accession number CRL 9800; and (b) recovering the transforming growth factor-β2 from the culture.
Claim 37
Independent Claim
36 words
A method for producing transforming growth factor-β2, comprising (a) culturing transfectant CHO-1/39, 12.5, CL 36 as deposited with the ATCC and assigned accession number CRL 9800; and (b) recovering the transforming growth factor-β2 from the culture.
Claim 38
Dependent Claim
17 words
The method according to claim 20 in which the transfectant is cultured in the presence of methotrexate.
Claim 39
Independent Claim
33 words
A method for producing transforming growth factor-β2, comprising (a) culturing transfectant CHO β2(414)c1.32 as deposited with the ATCC and assigned accession number ; and (b) recovering the transforming growth factor-02 from the culture.
Claim 40
Independent Claim
33 words
A method for producing transforming growth factor-β2, comprising (a) culturing transfectant CHO β2(414)c1.32 as deposited with the ATCC and assigned accession number ; and (b) recovering the transforming growth factor-02 from the culture.
Claim 41
Dependent Claim
17 words
The method according to claim 25 in which the transfectant is cultured in the presence of methotrexate.
Claim 42
Dependent Claim
33 words
A eucaryotic cell containing a nucleotide sequence encoding transforming growth factor-β2 under the control of a second nucleotide sequence that regulates gene expression so that the eucaryotic cell produces active transforming growth factor-β2.
Claim 43
Dependent Claim
33 words
A eucaryotic cell containing a nucleotide sequence encoding transforming growth factor-β2 under the control of a second nucleotide sequence that regulates gene expression so that the eucaryotic cell produces active transforming growth factor-β2.
Claim 44
Dependent Claim
33 words
The eucaryotic cell according to claim 27 in which the nucleotide sequence encoding the transforming growth factor-,82 comprises the nucleotide sequence substantially as depicted in FIG. 1 from nucleotide number -70 to 1755.
Claim 45
Dependent Claim
33 words
The eucaryotic cell according to claim 28 in which the nucleotide sequence encoding the transforming growth factor-82 comprises the nucleotide sequence substantially as depicted in FIG. 1a from nucleotide number -467 to 2111.
Claim 46
Dependent Claim
14 words
The eucaryotic cell according to claim 27 which comprises a Chinese Hamster Ovary cell.
Claim 47
Dependent Claim
12 words
The eucaryotic cell according to claim 27 which comprises a COS cell.
Claim 48
Dependent Claim
21 words
The eucaryotic cell according to claim 27 in which the second nucleotide sequence which controls gene expression comprises an SV40 promoter.
Claim 49
Dependent Claim
45 words
The eucaryotic cell according to claim 27 in which the second nucleotide sequence comprises a promoter and a sequence encoding a selectable marker for which the eucaryotic cell is deficient, so that eucaryotic cell containing the simian transforming growth factor-β2 coding sequence can be identified.
Claim 50
Dependent Claim
15 words
The eucaryotic cell according to claim 33 in which the selectable marker comprises dihydrofolate reductase.
Claim 51
Dependent Claim
16 words
The eucaryotic cell according to claim 34 which comprises a dihydrofolate reductase-deficient Chinese Hamster Ovary cell.
Claim 52
Dependent Claim
19 words
A cell line comprising CHO-1β9, 12.5, CL 36 as deposited with the ATCC and assigned accession number CRL 9800.
Claim 53
Dependent Claim
19 words
A cell line comprising CHO-1β9, 12.5, CL 36 as deposited with the ATCC and assigned accession number CRL 9800.
Claim 54
Dependent Claim
15 words
A cell line comprising CHO β2(414)c1.32 as deposited with the ATCC and assigned accession number
Claim 55
Dependent Claim
15 words
A cell line comprising CHO β2(414)c1.32 as deposited with the ATCC and assigned accession number
Claim 56
Dependent Claim
30 words
A substantially pure polypeptide comprising the amino acid sequence substantially as depicted in FIG. 1 b from about amino acid residue number 279 to about amino acid residue number 390.
Claim 57
Dependent Claim
30 words
A substantially pure polypeptide comprising the amino acid sequence substantially as depicted in FIG. 1 b from about amino acid residue number 279 to about amino acid residue number 390.
Claim 58
Independent Claim
19 words
A method for the treatment of tumors in vivo comprising administering transforming growth factor-β2 at a tumoricidal effective dose.
Claim 59
Independent Claim
19 words
A method for the treatment of tumors in vivo comprising administering transforming growth factor-β2 at a tumoricidal effective dose.
Claim 60
Independent Claim
23 words
A method for augmenting wound healing comprising administering a composition containing recombinant transforming growth factor-β2 at a concentration effective in stimulating cell proliferation.
Claim 61
Independent Claim
23 words
A method for augmenting wound healing comprising administering a composition containing recombinant transforming growth factor-β2 at a concentration effective in stimulating cell proliferation.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.