Claims for Patent: 8,969,064

Summary for Patent: 8,969,064

Title:	Gene expression technique
Abstract:	The present invention provides a method for producing a desired protein (such as a desired heterologous protein) comprising: (a) providing a host cell comprising a first recombinant gene encoding a protein comprising the sequence of a first chaperone protein, a second recombinant gene encoding a protein comprising the sequence of a second chaperone protein and a third gene, such as a third recombinant gene, encoding a desired protein (such as a desired heterologous protein), wherein the first and second chaperones are different; and (b) culturing the host cell in a culture medium to obtain expression of the first, second and third genes.
Inventor(s):	Finnis; Christopher John Arthur (Nottingham, GB), Sleep; Darrell (Nottingham, GB), Shuttleworth; Gillian (Nottingham, GB)
Assignee:	Novozymes Biopharma DK A/S (Bagsvaerd, DK)
Application Number:	13/474,313
Patent Claims:	1. A method for producing a desired protein comprising: (a) providing a host cell comprising a first recombinant gene encoding a protein comprising the sequence of protein disulphide isomerase, a second recombinant gene encoding a protein comprising the sequence of a JEM1 protein and a third gene encoding a desired protein; and (b) culturing the host cell in a culture medium to obtain expression of the first, second and third genes. 2. The method of claim 1 further comprising the step of purifying the thus expressed desired protein from the cultured host cell or the culture medium. 3. The method of claim 2 further comprising the step of lyophilising the purified protein. 4. The method of claim 2 further comprising the step of formulating the purified desired protein with a carrier or diluent and optionally presenting the thus formulated protein in a unit dosage form. 5. The method according to claim 1 wherein at least one of the first or second recombinant genes is encoded by a chromosomally integrated recombinant gene. 6. The method according to claim 1 wherein at least one of the first or second recombinant genes is encoded by a gene on a plasmid. 7. The method according to claim 1 wherein the third gene which encodes the desired protein is integrated in the chromosome of the host cell, or is provided on a plasmid within the host cell. 8. The method according to claim 6 wherein the plasmid is, or is not, a 2 .mu.m-family plasmid. 9. The method according to claim 8 wherein the plasmid comprises a gene encoding a protein comprising the sequence of a first chaperone protein and/or a gene encoding a protein comprising the sequence of a second chaperone protein, and a gene encoding a desired heterologous protein. 10. The method according to claim 8 wherein the plasmid is a disintegration vector. 11. The method according to claim 1 wherein the desired protein comprises a leader sequence effective to cause secretion from the host cell. 12. The method according to claim 1 wherein the desired protein is a eukaryotic protein, or a fragment or variant thereof, optionally a vertebrate or a fungal protein. 13. The method according to claim 1 wherein the desired protein is a commercially useful protein. 14. The method according to claim 1 wherein the desired protein comprises a sequence selected from the group consisting of albumin, a monoclonal antibody, an etoposide, a serum protein, antistasin, a tick anticoagulant peptide, transferrin, lactoferrin, endostatin, angiostatin, collagens, immunoglobulins, or immunoglobulin-based molecules or fragment of either, a Kunitz domain protein, interferons, interleukins, IL10, IL11, IL2, interferon .alpha. species and sub-species, interferon .beta. species and sub-species, interferon .gamma. species and sub-species, leptin, CNTF, CNTF.sub.Ax15, IL1-receptor antagonist, erythropoietin (EPO) and EPO mimics, thrombopoietin (TPO) and TPO mimics, prosaptide, cyanovirin-N, 5-helix, T20 peptide, T1249 peptide, HIV gp41, HIV gp120, urokinase, prourokinase, tPA, hirudin, platelet derived growth factor, parathyroid hormone, proinsulin, insulin, glucagon, glucagon-like peptides, insulin-like growth factor, calcitonin, growth hormone, transforming growth factor .beta., tumour necrosis factor, G-CSF, GM-CSF, M-CSF, FGF, coagulation factors in both pre and active forms, including but not limited to plasminogen, fibrinogen, thrombin, pre-thrombin, pro-thrombin, von Willebrand's factor, .alpha..sub.1-antitrypsin, plasminogen activators, nerve growth factor, LACI, platelet-derived endothelial cell growth factor (PD-ECGF), glucose oxidase, serum cholinesterase, aprotinin, amyloid precursor protein, inter-alpha trypsin inhibitor, antithrombin III, apo-lipoprotein species, Protein C, Protein S, a metabolite, antibiotic, and a variant or fragment of these. 15. The method according to claim 1 wherein the desired protein comprises the sequence of albumin or a variant or fragment thereof. 16. The method according to claim 1 wherein the desired protein comprises the sequence of a transferrin family member, optionally transferrin or lactoferrin, or a variant or fragment thereof. 17. The method according to claim 1 wherein the desired protein is a desired heterologous protein that comprises a fusion protein, a fusion protein of albumin or a transferrin family member or a variant or fragment of either, fused directly or indirectly to the sequence of another protein.

Details for Patent 8,969,064

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Bayer Healthcare Pharmaceuticals Inc.	TRASYLOL	aprotinin	Injection	020304	12/29/1993		2023-12-23
Microbix Biosystems Inc.	KINLYTIC	urokinase	For Injection	021846	01/16/1978		2023-12-23
Nps Pharmaceuticals, Inc.	NATPARA	parathyroid hormone	For Injection	125511	01/23/2015		2023-12-23