Integrated continuous manufacturing of therapeutic protein drug substances

Patent metadata

Field	Value
Publication number	US-9650413-B2
Application number	US-201514627559-A
Country	US
Kind code	B2
Filing date	Feb 20, 2015
Priority date	Mar 8, 2013
Publication date	May 16, 2017
Grant date	May 16, 2017

Abstract

Provided herein are integrated continuous biomanufacturing processes for producing a therapeutic protein drug substance. Also provided are systems that are capable of continuously producing a therapeutic protein drug substance.

First claim

What is claimed is:

1. An integrated and continuous process for manufacturing a therapeutic protein drug substance, the process comprising:
(i) culturing recombinant therapeutic protein-secreting cells in a perfusion bioreactor comprising a liquid culture medium under conditions that allow the cells to secrete the recombinant therapeutic protein into the medium, and wherein at least 90% cell-free volumes of the medium are continuously or periodically removed from the perfusion bioreactor and fed into a first periodic counter current chromatography system (PCCS1);
(ii) capturing the recombinant therapeutic protein from the medium using the PCCS1, wherein an eluate of the PCCS1 comprising the recombinant therapeutic protein is continuously fed into a second periodic counter current chromatography system (PCCS2); and
(iii) purifying and polishing the recombinant therapeutic protein using the PCCS2, wherein the purifying is performed using a resin in the PCCS2 that is different in chemical structure compared to the resin in the PCCS2 used to perform the polishing, and an eluate from the PCCS2 is the therapeutic protein drug substance; and
wherein the process is integrated and runs continuously from the culturing step to the purifying and polishing step, and the elapsed time between feeding the medium into the PCCS1 and eluting the therapeutic protein drug substance from the PCCS2 is about 4 hours to about 9 hours.

2. The process of claim 1 , wherein the PCCS1 performs at least two different unit operations.

3. The process of claim 2 , wherein the use of the PCCS1 or the PCCS2, or both, includes column switching.

4. The process of claim 2 , wherein the PCCS1 performs the unit operations of capturing the recombinant therapeutic protein and inactivating viruses.

5. The process of claim 1 , wherein the PCCS1 or the PCCS2, or both, utilizes at least two chromatography columns.

6. The process of claim 1 , wherein the PCCS1 comprises four chromatography columns, wherein three of the four chromatography columns in the PCCS1 perform the unit operation of capturing the recombinant therapeutic protein from the medium.

7. The process of claim 6 , wherein the capturing is performed using affinity chromatography, cation exchange chromatography, anion exchange chromatography, or molecular sieve chromatography.

8. The process of claim 7 , wherein the affinity chromatography is performed with a capture mechanism selected from the group consisting of: protein A-binding capture mechanism, substrate-binding capture mechanism, antibody-or antibody fragment-binding capture mechanism, aptamer-binding capture mechanism, and cofactor-binding capture mechanism.

9. The process of claim 8 , wherein the affinity chromatography is performed with a protein A-binding capture mechanism, and the recombinant therapeutic protein is an antibody or an antibody fragment.

10. The process of claim 6 , wherein an eluate comprising the recombinant therapeutic protein from the three of the four chromatography columns in the PCCS1 is fed into the fourth column of the PCCS1.

11. The process of claim 10 , wherein the fourth chromatography column of the PCCS1 performs the unit operation of inactivating viruses by incubating the eluate from the three of the four chromatography columns in the PCCS1 at a pH of about 3.0 to 5.0.

12. The process of claim 11 , further comprising adjusting the pH of an eluate comprising the recombinant therapeutic protein from the fourth chromatography column of the PCCS1 using an in-line buffer adjustment reservoir before the eluate from the fourth column of the PCCS1 is fed into the PCCS2.

13. The process of claim 12 , wherein the PCCS2 comprises three chromatography columns and a chromatographic membrane, wherein the three chromatography columns in the PCCS2 perform the unit operations of purifying the recombinant therapeutic protein from the eluate from the fourth chromatography column of the PCCS1 through cation or anion exchange chromatography, and an eluate comprising the recombinant protein from the three chromatography columns in the PCCS2, is fed into the chromatographic membrane in the PCCS2.

14. The process of claim 13 , wherein the chromatographic membrane in the PCCS2 performs the unit function of polishing the recombinant therapeutic protein present in the eluate from the three chromatography columns in the PCCS2 through cation or anion exchange chromatography.

15. The process of claim 14 , wherein the chromatographic membrane in the PCCS2 performs the unit function of polishing through cation exchange chromatography.

16. The process of claim 1 , further comprising formulating the therapeutic protein drug substance into a pharmaceutical composition.

17. The process of claim 1 , wherein the recombinant therapeutic protein is an antibody or antibody fragment, an enzyme, an engineered protein, or an immunogenic protein or protein fragment.

18. The process of claim 1 , wherein the entire process does not include storing the recombinant therapeutic protein in a break tank.

19. The process of claim 1 , wherein the entire process includes storing the recombinant therapeutic protein in 1, 2, or 3 break tank(s).

20. The process of claim 19 , wherein the 1, 2, or 3 break tank(s) are used to store the therapeutic protein for a total time period of about 5 minutes to about 4 hours.

21. The process of claim 19 , wherein the 1, 2, or 3 break tank(s) has/have a volume of 1 mL to about 100 mL.

22. The method of claim 1 , wherein the elapsed time between feeding the medium into the PCCS1 and eluting the therapeutic protein drug substance from the PCCS2 is greater than about 4 hours and is less than about 7 hours.

23. The method of claim 22 , wherein the elapsed time between feeding the medium into the PCCS1 and eluting the therapeutic protein drug substance from the PCCS2 is greater than about 4 hours and is less than about 6 hours.

24. The method of claim 23 , wherein the elapsed time between feeding the medium into the PCCS1 and eluting the therapeutic protein drug substance from the PCCS2 is greater than about 4 hours and is less than about 5 hours.

25. An integrated and continuous process for manufacturing a therapeutic protein drug substance, the process comprising:
(i) culturing recombinant therapeutic protein-secreting mammalian cells in a perfusion bioreactor comprising a liquid culture medium under conditions that allow the cells to secrete the recombinant therapeutic protein into the medium, and wherein at least 90% cell-free volumes of the medium are continuously or periodically removed from the perfusion bioreactor and fed from the perfusion bioreactor into a first periodic counter current chromatography system (PCCS1);
(ii) capturing the recombinant therapeutic protein from the medium using the PCCS1, wherein an eluate of the PCCS1 comprising the recombinant therapeutic protein is continuously fed into a second periodic counter current chromatography system (PCCS2); and
(iii) purifying and polishing the therapeutic recombinant protein using the PCCS2, wherein the purifying is performed using a resin in the PCCS2 that is different in chemical structure compared to the resin in the PCCS2 used to perform the polishing, and an eluate from the PCCS2 is the therapeutic protein drug substance;
wherein:
the process is integrated and runs continuously from the culturing step to the purifying and polishing step,
the entire process includes stori

Assignees

• Genzyme Corp

Inventors

• Konstantinov Konstantin
• Godawat Rahul
• Warikoo Veena
• Jain Sujit

Classifications

• C12N9/2402

  hydrolysing O- and S- glycosyl compounds (3.2.1) · CPC title

• B01D15/422

  Displacement mode · CPC title

• C12Y302/01022

  Alpha-galactosidase (3.2.1.22) · CPC title

• B01D15/327

  with hydrophobic interaction · CPC title

• C12Y302/01045

  Glucosylceramidase (3.2.1.45), i.e. beta-glucocerebrosidase · CPC title