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Last Updated: April 27, 2024

Claims for Patent: 9,914,770

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Summary for Patent: 9,914,770

Title:	Cloning, expression and purification method for the preparation of ranibizumab
Abstract:	A polynucleotide sequence and a method for Ranibizumab cloning, expression and production having better yield and biologically active protein.
Inventor(s):	Shandilya; Harish (Gujarat, IN), Gadgil; Himanshu (Gujarat, IN), Farkade; Vivek (Gujarat, IN)
Assignee:	INTAS Pharmaceuticals Ltd (Ahmedabad, IN)
Application Number:	14/787,981
Patent Claims:	1. A process for the preparation of Ranibizumab comprising steps of: a) transforming a first host cell with a first vector comprising a first polynucleotide encoding a signal sequence of SEQ ID NO: 5 operably linked to a light chain of Ranibizumab of SEQ ID NO: 2, wherein the first polynucleotide is operably linked to an inducible promoter system; b) transforming a second host cell with a second vector comprising a second polynucleotide encoding a signal sequence of SEQ ID NO: 6 operably linked to a heavy chain of Ranibizumab of SEQ ID NO: 4, for heavy chain of wherein the second polynucleotide is operably linked to an inducible promoter system; c) separately culturing the first host cell and the second host cell in a growth medium; d) expressing the light chain of Ranibizumab in the first host cell and the heavy chain of Ranibizumab in the second host cell as periplasmic inclusion bodies; e) solubilizing the inclusion bodies; and f) refolding in-vitro the solubilized light chain and heavy chain of Ranibizumab. 2. The process of claim 1, wherein the signal sequence of SEQ ID NO: 5 in the expressing step directs the first host cell to transport the light chain of Ranibizumab to a periplasmic space of the first host cell. 3. The process of claim 1, wherein the signal sequence of SEQ ID NO: 6 in the expressing step directs the second host cell to transport the heavy chain of Ranibizumab to a periplasmic space of the second host cell. 4. The process of claim 1, wherein the first host cell in the culturing step is cultured to an OD600 of about 50. 5. The process of claim 1, wherein the second host cell in the culturing step is cultured to an OD600 of about 100. 6. The process of claim 1, wherein the first polynucleotide is nt 8 to nt 718 of SEQ ID NO: 1. 7. The process of claim 1, wherein the second polynucleotide is nt 9 to nt 770 of SEQ ID NO: 3.

Details for Patent 9,914,770

Glossary

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Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	LUCENTIS	ranibizumab	Injection	125156	06/30/2006	⤷ Try a Trial	2033-04-30
Genentech, Inc.	LUCENTIS	ranibizumab	Injection	125156	08/10/2012	⤷ Try a Trial	2033-04-30
Genentech, Inc.	LUCENTIS	ranibizumab	Injection	125156	10/13/2016	⤷ Try a Trial	2033-04-30
Genentech, Inc.	LUCENTIS	ranibizumab	Injection	125156	03/20/2018	⤷ Try a Trial	2033-04-30
>Applicant	>Tradename	>Biologic Ingredient	>Dosage Form	>BLA	>Approval Date	>Patent No.	>Expiredate

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