Removal of residual cell culture impurities

Inactive Publication Date: 2016-10-06
NOVARTIS AG
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  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0008]The invention thus includes methods, which increase the yield, purity and/or safety of biological products produced from cell culture. Biological products prepared by methods of the invention may include, but are not limited to: biopharmaceuticals, proteins, polysaccharides, viral antigens, and antibodies. In a particular aspect, the method provides a biological product substantially free of residual DNA.
[0009]The inventors have surprisingly found that purification of sample comprising protein and DNA derived from cell culture is significantly improved by an addition of an anionic detergent to a solution comprising the proteins and cellular DNA, followed by a purification step comprising an ion exchange matrix. The problem to be solved might relate to the inefficiency of separation of negatively charged DNA impurities from proteins of interest on a positively charged ion exchange matrix. It can be assumed that secondary interactions were playing a role in the dimi

Problems solved by technology

The problem to be solved might relate to the inefficiency of separation of negatively cha

Method used

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  • Removal of residual cell culture impurities
  • Removal of residual cell culture impurities
  • Removal of residual cell culture impurities

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Example

2. The method of embodiment 1, wherein the first solution is selected from the group consisting of: cell or tissue lysates, culture media, cell culture supernatants, plasma, and partially purified protein solutions.

3. The method of any one of the preceding embodiments, wherein the protein of interest is selected from the group consisting of: therapeutic proteins, immunogenic proteins (e.g., viral antigens), and antibodies or antigen-binding fragments thereof.

4. The method of any one of the preceding embodiments, wherein the anionic detergent is selected from the group consisting of: fatty acid detergents.

5. The method of any one of the preceding embodiments, wherein the anionic detergent is different from any other detergent used in a process of protein purification.

6. The method of any one of the preceding embodiments, wherein the anionic detergent does not include deoxycholate and / or sodium lauryl sulfate.

7. The method of any one of the preceding embodiments, wherein the ion excha...

Example

14. The method of embodiment 12 or 13, wherein the pharmaceutical composition is a prophylactic composition, therapeutic composition, or combination thereof.

15. The method of any one of embodiments 12-14, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

16. The method of any one of embodiments 12-15, wherein the pharmaceutical composition further comprises and adjuvant.

17. The method of any one of embodiments 12-16, further comprising a step of packaging the pharmaceutical composition into a sterile closed system.

Example

18. The method of embodiment 17, wherein the sterile closed system is selected from the group consisting of: vials, syringes, and containers.

19. The method of embodiment 17 or 18, wherein the sterile closed system is plastic or glass.

20. The method of any one of embodiments 17-19, wherein the sterile closed system comprises a siliconized surface.

21. A use of the pharmaceutical composition of any one of embodiments 12-20, for the manufacture of a medicament for administering a subject in need thereof.

22. The pharmaceutical composition of any one of embodiments 12-20 for use as a medicament for administering to a subject.

23. A method comprising administering to a subject the pharmaceutical composition of any one of embodiments 12-20.

24. A viral vaccine comprising no more than 5 ng of residual DNA and no more than 1.0 μg nucleoprotein per dose.

25. The viral vaccine of embodiment 24, comprising no more than 1 ng of residual DNA and no more than 0.5 μg nucleoprotein per dose.

26. The vira...

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Abstract

The present application discloses methods for removing residual impurities from protein preparations. Such methods include addition of an anionic detergent to a solution comprising proteins of interest and cellular contaminants under non-precipitating conditions and passing the solution through an ion exchange column.

Description

RELATED APPLICATIONS[0001]This international patent application claims priority to U.S. Provisional Application 61 / 904,747 filed Nov. 15, 2013 and European Application 13199257.0 filed Dec. 20, 2013, the contents of each of which are hereby incorporated by reference in its entirety.FIELD OF THE INVENTION[0002]This invention relates to production of proteins in host cells and improved purification methods thereof.BACKGROUND OF THE INVENTION[0003]Various methods for producing vaccines and other biologics in cell cultures have been pre-described. If continuous cell lines are used for the production, there is the risk that residual DNA of the cell line could be oncogenic. It is therefore required to destroy and remove residual DNA from therapeutic proteins of interests. For viral vaccines the FDA currently recommends a DNA amount of less than 10 ng / dose and a fragment size of less than 200 base pairs (Guidance of Industry. Characterization and Qualification of Cell Substrates and other ...

Claims

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Application Information

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IPC IPC(8): A61K39/145C12N7/00
CPCA61K39/145C12N7/00A61K2039/53C12N2760/16151C12N2760/16134C07K1/18A61K39/12A61P31/16A61P37/04B01D15/361C07K14/005
Inventor NORMAN, CARNLEYSUDA, ERICDOWLESS, KAYLAASTIGARRAGA, RUIZBASTEK, PATRICKYANNONE, VAISHALI
Owner NOVARTIS AG
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