Multicellular living organisms and unmodified parts thereof and – Method of using a transgenic nonhuman animal to manufacture... – The protein is isolated or extracted from milk
Reexamination Certificate
2000-07-28
2002-10-29
Crouch, Deborah (Department: 1632)
Multicellular living organisms and unmodified parts thereof and
Method of using a transgenic nonhuman animal to manufacture...
The protein is isolated or extracted from milk
C800S014000, C435S320100, C436S023000
Reexamination Certificate
active
06472584
ABSTRACT:
FIELD OF INVENTION
The present inventions involves the large-scale production of membrane specific proteins and more particularly involves the production of such proteins using transgenic animals and most specifically involves the production of CFTR, the protein involved with Cystic Fibrosis.
BACKGROUND OF THE INVENTION
Striking progress has been made in our understanding of cystic fibrosis (CF) during the period since the gene associated with the disease was identified
1-3
. Many mutations within the gene have been identified in DNA from patients with CF
4-10
. The protein product of the gene, named CFTR, has been identified
11
and functional studies have shown that CFTR cDNA is able to complement the defect in ion transport characteristic of cells from CF patients
12,13
. The domain structure of CFTR has been probed by analysis of mutated versions of the protein
14-18
. Such studies indicate that CFTR is a Cl
−
channel
14,19,20
and that it is regulated by phosphorylation of the R-domain
16,18,21
and by nucleotide binding
22
. To our knowledge none of such proteins have involved exogenous or recombinant membrane proteins.
SUMMARY OF THE INVENTION
In accordance with the principles of the present invention it has been surprisingly discovered that exogenous or recombinant membrane proteins can be produced in the milk of transgenic animals. In the most preferred embodiment of the present invention, recombinant CFTR has been produced in the milk of transgenic mice containing CFTR cDNA downstream of a mammary specific promoter. Most preferred embodiments will employ larger transgenic animals including, for example, rabbits, goats, sheep and cows which produce large quantities of milk however, the development of such animals takes considerably longer than the time periods required for developing transgenic mice. While the time periods vary considerably, the procedures and methods are substantially identical.
Other embodiments include the apocrine like secretion in tissue culture of membrane proteins by cells which have been maintained in a differentiated state. Such proteins would be secreted as part of the membrane released during the “pinching off” proces. Membrane proteins suitable for production according to the methods of the present include receptors, channels, viral glycoproteins, transporters and other proteins typically associated with cellular membranes. Such membrane proteins can be used for therapeutics such as by administration of the abnormally deficient or missing protein, or by use as vaccines in the case of viral glycoproteins including for example utilization of the envelope of HIV, Herpes Virus or influenza in order to develop immunity with respect to infection caused thereby.
Advantageously, methods of the present invention can also be employed for the production of membrane proteins useful for diagnostic purposes. For example, one could use the present methods to produce the receptor for thyroid stimulating hormone (TSHR) which would be useful for the diagnosis of Graves Disease. Such proteins could also be used to screen for therapeutically active compounds.
Preferred methods of the present invention will employ promoters, ideally coupled with suitable enhancers, that are mammary specific so that production of the desired membrane protein is incorporated into milk fat globules which will appear in the milk of a lactating female transgenic animal. Such an animal is, of course, by definition, reproductively competant. Greater appreciation of these and other emodiments will be acquired by study of the following drawings, examples and detailed procedures.
REFERENCES:
patent: 4873316 (1989-10-01), Meade et al.
patent: 5240846 (1993-08-01), Collins et al.
patent: 5304489 (1994-04-01), Rosen
patent: 5322775 (1994-06-01), Clark et al.
patent: 5574206 (1996-11-01), Jolicoeur
patent: 5589604 (1996-12-01), Drohan et al.
patent: 5633076 (1997-05-01), Deboer
patent: 0 264 166 (1988-04-01), None
patent: 0 446 017 (1991-09-01), None
Anderson, M. et al., (1991) “Generation of cAMP-Activated Chloride Currents by Expression of CFTR”,Science, 251:679-682.
Andres, A.C. et al., (1987) “Ha-ras Oncogene Expression Directed By a Milk Protein Gene Promoter: Tissue Specificity, Hormonal Regulation, and Tumor Induction in Transgenic Mice”Proc. Natl. Acad. Science, USA, 84:1299-1303.
Bagley, R.G. et al., (1994) “Delivery of Purified, Functional CFTR to Epithelial Cells In Vitro Using Influenza Hemagglutinin” in Late-Breaking Science Platform Presentations and Posters, Eighth Annual North American Cystic Fibrosis Conference (abstract).
Buhler, Th. Et al., (1990) “Rabbit &bgr;-Casein Promoter Directs Secretion of Human Interleukin-2 Into the Milk of Transgenic Rabbits”Bio/Technology,8:140-143.
Clark et al.,Bio/Technology, vol. 7 (May 1989) pp. 487-492.
Clark, A.J. et al., (1987) “Pharmaceuticals from transgenic livestock”Tibtech, 5:20-24.
Clark, A.J. et al., (1989) “Expression of Human Anti-Hemophilic Factor IX in the Milk of Transgenic Sheep”Bio/Technology, 7:487-492.
Cosman et al.,Mol Immunol.vol. 23, No. 9 (1986) pp. 935-941.
Denman, J. et al. (1991) “Transgenic Expression of a Variant of Human Tissue-Type Plasminogen Activator in Goat Milk: Purification and Characterization of the Recombinant Enzyme”Bio/Technology, 9:839-843.
DiTullio, P. et al. (1992) “Production of Cystic Fibrosis Transmembrane Conductance Regulator in the Milk of Transgenic Mice”Bio/Technology,10:74-77.
Ebert, K.M. et al., (1991) “Transgenic Expression of a Variant of Human Tissue-Type Plasminogen Activator in Goat Milk: Generation of Transgenic Goats and Analysis of Expression”Bio/Technology, 9:835-838.
Gordon, K. et al., (1987) “Production of Human Tissue Plasminogen Activator in Transgenic Mouse Milk”Bio/Technology,5:1183-1187.
Gregory, R. et al., (1990) “Expression and characterization of the cystic fibrosis transmembrane conductance regulator”Nature, 347(6291):382-386.
Harris et al., “Gene expression in the mammary gland”,J. Reprod. Fert.(1990) 88:707-715.
Hennighausen, L.G. (1982) “Characterization and Cloning of the mRNAs Specific for the Lactating Mouse Mammary Gland”Eur. J. Biochem,125:131-141.
Houdebine, Louis-Marie, “Production of Pharmaceutical Proteins from Transgenic Animals”Journal of Biotechnology,vol. 34, pp. 269-287.
Imam, A. et al., (1981) “Isolation and Characterization of a Major Glycoprotein From Milk-Fat-Globule Membrane of Human Breast Milk”Biochem. J.,193:47-54.
Jolicoeur et al., “Efficient production of human immunodeficiency virus proteins in transgenic mice”Journal of Virology, vol. 66, No. 6, pp. 3904-3908.
Kartner, N. et al., (1991) “Expression of the Cystic Fibrosis Gene in Non-Epithelial Invertebrate Cells Produces a Regulated Anion Conductance”Cell, 64-681-691.
Kerem, B. S. et al., (1989) “Identification of the Cystic Fibrosis Gene: Genetic Analysis”Science, 245:1073-1080.
Krimpenfort et al., “Generation of Transgenic Dairy Cattle Using ‘In Vitro’ Embryo Production”Biotechnology,vol. 9, pp. 844-847.
Meade, H. et al., (1990) “Bovine Alphas1-Casein Gene Sequences Direct High Level of Expression of Active Human Urokinase in Mouse Milk”Bio/Technology, 8:443-446.
Mullins et al., “Transgenesis in Nonmurine Species”Hypertension, vol. 22, No. 4, pp. 630-633.
Patton, S. et al., (1986) “A Method for Isolation of Milk Fat Globules”Lipids, 21(2):170-174.
Patton S. et al., (1975) “The Milk Fat Globule Membrane”Biochem. Biophys. Acta, 415:273-309.
Rich et al.,Nature, vol. 347 (Sep. 27, 1990) pp. 358-363.
Riordan, J. et al., (1989) “Identification of the Cystic Fibrosis Gene: Cloning, and Characterization of Complementary DNA”Science, 245:1066-1073.
Rommens, J.M. et al., (1989) “Identification of the Cystic Fibrosis Gene: Chromosome Walking and Jumping”Science, 245:1059-1065.
Sasaki, M. et al., (1978) “Lactose and Major Milk Proteins are Present in Secretory Vesicle-Rich Fractions From Lactating mammary Gland”Proc. Natl. Acad. Sci. USA, 75(10):5020-5024.
Suda et al., “Induction of a variety of tumors by c-erbB2 and clonal nature of lymphomas even with the mutated gene (Val659 Glu569)”The
Fish & Richardson PC
Genzyme Corporation
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