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Last Updated: April 24, 2024

Claims for Patent: 9,163,257

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Summary for Patent: 9,163,257

Title:	Expression vector and methods of producing high levels of proteins
Abstract:	A process for high expression of protein of interest using an expression vector. The process comprises at least the following regulatory elements: a) a CMV promoter, or its functional variants, b) an intron, c) TPL or its functional variants, d) VA genes or functional variants, and e) a bovine growth hormone polyadenylation sequence or functional variants.
Inventor(s):	Singh; Arun K. (Gujarat, IN), Goel; Ashish (Gujarat, IN), Mendiratta; Sanjeev K. (Gujarat, IN)
Assignee:	Cadila Healthcare Limited (Ahmedabad, IN)
Application Number:	11/922,548
Patent Claims:	1. A mammalian expression vector comprising: (i) a CMV promoter; (ii) an intron consisting of the nucleic acid sequence of SEQ ID NO: 1, (iii) a tripartite leader (TPL); (iv) a virus associated (VA) gene; (v) a bovine growth hormone (BGH) polyadenylation sequence; and (vi) a nucleic acid sequence encoding a protein of interest. 2. The vector of claim 1, wherein the TPL consists of the nucleic acid sequence of SEQ ID NO: 4. 3. The vector of claim 1, wherein the VA gene consists of the nucleic acid sequence of SEQ ID NO: 5. 4. The vector of claim 1, further comprising a selection marker selected from the group consisting of dihydrofolate reductase, adenosine deaminase, ornithine decarboxylase, asparagine synthetase, and glutamine synthetase. 5. The vector of claim 1, wherein the protein is recombinant erythropoietin. 6. The vector of claim 1, wherein the protein is TNFR-IgGFc. 7. The vector of claim 1, wherein the protein is rituximab, trastuzumab, bevacuzumab or other monoclonal antibody. 8. An isolated mammalian cell comprising the vector of claim 1. 9. The isolated mammalian cell of claim 8, wherein the cell is selected from the group consisting of a Cos, CHO, CHO DHFR-, BHK1, and NS0 cell. 10. A mammalian expression vector comprising: (i) a CMV promoter; (ii) an intron consisting of the nucleic acid sequence of SEQ ID NO: 1, (iii) a tripartite leader (TPL) consisting of the nucleic acid sequence of SEQ ID NO: 4; (iv) a virus associated (VA) gene consisting of the nucleic acid sequence of SEQ ID NO: 5; (v) a bovine growth hormone (BGH) polyadenylation sequence; and (vi) a nucleic acid sequence encoding a protein of interest. 11. The vector of claim 10, wherein the protein is recombinant erythropoietin. 12. The vector of claim 10, wherein the protein is TNFR-IgGFc. 13. An isolated mammalian cell comprising the vector of claim 10. 14. The vector of claim 10, further comprising a selection marker selected from the group consisting of dihydrofolate reductase, adenosine deaminase, ornithine decarboxylase, asparagine synthetase, and glutamine synthetase. 15. A method for expressing a protein in CHO cells comprising: (a) constructing a vector comprising: (i) a CMV promoter; (ii) an intron consisting of the nucleic acid sequence of SEQ ID NO: 1, (iii) a tripartite leader (TPL); (iv) a virus associated (VA) gene; (v) a bovine growth hormone (BGH) polyadenylation sequence; and (vi) a nucleic acid sequence encoding a protein of interest; and (b) transfecting CHO cells with the vector such that the protein is expressed. 16. The method of claim 15, wherein the protein is recombinant erythropoietin. 17. The method of claim 15, wherein the protein is TNFR-IgGFc. 18. The method of claim 15, wherein the protein is rituximab, trastuzumab, bevacuzumab or other monoclonal antibody. 19. The method of claim 15, wherein the protein is selected from the group consisting of TN FR-IgGFc, rituximab, trastuzumab, bevacuzumab or other monoclonal antibody.

Details for Patent 9,163,257

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Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986	⤷ Try a Trial	2025-06-20
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132		⤷ Try a Trial	2025-06-20
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132		⤷ Try a Trial	2025-06-20
Genentech, Inc.	RITUXAN	rituximab	Injection	103705	11/26/1997	⤷ Try a Trial	2025-06-20
>Applicant	>Tradename	>Biologic Ingredient	>Dosage Form	>BLA	>Approval Date	>Patent No.	>Expiredate

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