Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Lymphokines – e.g. – interferons – interlukins – etc.
Patent
1992-10-08
1998-12-01
Budens, Robert D.
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Lymphokines, e.g., interferons, interlukins, etc.
424 851, 4241981, 530380, 530386, 530399, C07K 14495, A61K 3818, A61K 3819
Patent
active
058440852
ABSTRACT:
Recombinant transforming growth factor-.beta.1 (TGF-.beta.1) is expressed to high levels in Chinese hamster ovary (CHO) cells using dihydrofolate reductase (dhfr) gene amplification. The expression plasmid was derived from the pSV2 vectors and contained, in tandem, the simian TGF-.beta.1 and mouse dhfr cDNAs. Transcription of both cDNAs was controlled by the SV40 early promoter. Stepwise selection of transfected CHO cells in increasing concentrations of methotrexate yielded cell lines expressing amplified TGF-.beta.1 nucleic acid sequences. The expression plasmid DNA was amplified greater than 35-fold in one of the methotrexate selected transfectants. The major proteins secreted by these cells consisted of latent TGF-.beta.1 and TGF-.beta.1 precursor polypeptides as judged by immunoblots using site-specific anti-peptide antibodies derived from various regions of the TGF-.beta.1 precursor. Levels of recombinant TGF-.beta.1 protein secreted by these cells approached 30 ug/24 hour/10.sup.7 cells. Expression vectors encoding TGF-.beta.1 precursor variants which direct the synthesis and secretion of TGF-.beta.1 in transfected mammalian cells are also described.
REFERENCES:
patent: 4714613 (1987-12-01), Shouval et al.
patent: 4886747 (1989-12-01), Derynck et al.
Tucker et al., "Growth Inhibitor from BSC-1 Cells Closely Related to Platelet Type .beta.Transforming Growth Factor," Science 226:705-707, 9 Nov. 1984.
Derynck, et al., "Human Transforming Growth Factor-.beta. Complementary DNA Sequence and Expression in Normal and Transformed Cells," Nature 316:701-705, 22 Aug. 1985.
Roberts et al. "Type .beta. Transforming Growth Factor:A Bifunctional Regulator of Cellular Growth," Proc. Natl. Acad. Sci. USA 82:119-123, Jan., 1985.
Derynck et al., "The Murine Transforming Growth Factor-.beta. Precursor," J. Biol. Chem. 261(10):4377-4379, 1986.
Sharples et al., "Cloning and Sequence Analysis of Simian Transforming Growth Factor-.beta. cDNA," DNA 6(3):239-244, Jun. 1987.
Brunner Amy M.
Gentry Larry
Purchio Anthony F.
Twardzik Daniel
Bristol-Myers Squibb Pharmaceutical Research Institute-Seattle
Budens Robert D.
LandOfFree
Cloning and expression of simian transforming growth factor .bet does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Cloning and expression of simian transforming growth factor .bet, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Cloning and expression of simian transforming growth factor .bet will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2396596