Device and method for the production of biologically active compounds by fermentation

• US 20080187967A1
• Filed: 12/13/2007
• Published: 08/07/2008
• Est. Priority Date: 06/17/2005
• Status: Active Grant

• Current Assignee: Merz Pharma GmbH & Company Kgaa
• Original Assignee: Merz Pharma GmbH & Company Kgaa
• Inventors: Pfeil, Michael, Doelle, Bernd
• Granted Patent: US 7,927,836 B2
• Time in Patent Office: Days
• US Class Current: 435/71.3
• CPC Class Codes: C07K 14/33 from Clostridium (G)
• CPC Class Codes: C12M 37/02 Filters
• CPC Class Codes: C12M 41/14 Incubators; Climatic chambers per se B01L1/00
• CPC Class Codes: C12M 41/40 of pressure
• CPC Class Codes: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione
• CPC Class Codes: Y10S 435/809 Incubators or racks or holders for culture plates or containers

Abstract

The present invention relates to a process and an apparatus for the fermentational production of biologically active materials, wherein a fermenter is located in an insulator which, in turn, is located within a working chamber or is adjacent to it. A pressure gradient in relation to ambient pressure prevails in both the insulator and in the working chamber.

Claims

1. An apparatus for the fermentative production of a biologically active compound, comprising at least a first insulator containing a fermenter, the first insulator being surrounded or adjacent to a working chamber, wherein the working chamber is connected to the environment via a pressure sluice, wherein a low pressure prevails in the insulator and in the working chamber, wherein the pressure (in relation to ambient pressure) in the insulator is lower than the pressure (in relation to ambient pressure) in the working chamber, and wherein an over pressure prevails in the pressure sluice in relation to ambient pressure.

2. The apparatus according to claim 1, wherein the pressure in the insulator is 20 to 200 Pa lower than ambient pressure.

3. The apparatus according to claim 1, wherein the pressure in the working chamber of 5 to 50 Pa is lower than ambient pressure.

4. The apparatus according to claim 1, wherein the pressure in the pressure sluice of 10 to 100 Pa is higher than ambient pressure.

5. The apparatus according to claim 1, wherein the apparatus comprises at least a second insulator, and wherein the second insulator does not contain a fermenter.

6. The apparatus according to claim 1, wherein the apparatus and/or the first insulator and/or the second insulator comprise at least a supply air duct and an exhaust air duct, wherein the supply air duct and the exhaust air duct comprise a filter, selected from a HEPA filter.

7. The apparatus according to claim 5, wherein the internal pressure of the first insulator is equal to the internal pressure of the second insulator.

8. The apparatus according to claim 5, wherein the internal pressure of the first insulator is different than the internal pressure of the second insulator.

9. The apparatus according to claim 5, wherein the first and the second insulator are connected with each other via a passage, wherein the passage allows for material transport from the first insulator to the second insulator and also allows for material transport from the second insulator to the first insulator.

10. The apparatus according to claim 9, wherein the passage is a sluice.

11. The apparatus according claim 1, wherein the apparatus comprises a sterilizing facility and/or a disinfection facility.

12. The apparatus according to claim 1, wherein the first insulator comprises an anaerobic working fermenter.

13. The apparatus according to claim 1, wherein the first insulator comprises a precipitation facility.

14. The apparatus according to claim 1, wherein the first insulator comprises a filtration facility.

15. The apparatus according to claim 5, wherein the second insulator comprises an extraction facility.

16. The apparatus according to claims 5, wherein the second insulator comprises a precipitation facility.

17. The apparatus according to claim 5, wherein the second insulator comprises at least a chromatography facility.

18. A process for the fermentational production of a biologically active compound comprising:

a fermentation step for the production of the biologically active compound,

a purification step for the biologically active compound,

wherein the process is conducted in the apparatus according to claim 1.

19. The process according to claim 18, wherein the biologically active compound is a toxin or another protein obtained from fermentation, selected from a botulinum toxin and a botulinum neurotoxin.

20. The process according to claim 19, wherein the botulinum toxin is a botulinum toxin belonging to Clostridium botulinum of types A, B, C, D, E, F, and G, or is a mixture of two or more of these types.

21. The process according to claim 20, wherein the botulinum toxin or the mixture of botulinum toxins is/are a botulinum neurotoxin or a composition of botulinum neurotoxins.

22. The process according to claim 18, wherein the fermentation step occurs in the first insulator and the purification step occurs totally or partially in a second insulator.

23. The process according to claim 18, wherein the fermentation step and a part of the purification step occur in the first insulator and another part of the purification step occurs in the second insulator.

24. The process according to claim 18, wherein the fermentation step, a precipitation and filtration of a product of the fermentation step, occur in the first insulator as a part of the purification step.

25. The process according to claim 18, wherein the first insulator and the second insulator are operated at the same temperature or at different temperatures.

26. The process according to claim 18, wherein the temperature in the first and/or the second insulator are changed according to each process step.

27. The process according to claim 26, wherein the temperature in the first insulator is in a range of approximately 20°

to approximately 50°

C. and the temperature in the second insulator is in a range of approximately −

5°

to approximately +25°

C.

28. The process according to claim 18, wherein the process comprises the following steps:

a) in the first insulator

inoculating fermentation media with a producing strain which produces a biologically active compound;

fermenting the producing strain;

separating supernatant from cells of the producing strain; and

precipitating the supernatant;

b) in the second insulator

centrifuging the precipitated supernatant to obtain a pellet;

extracting the pellet, centrifuging to obtain a supernatant;

precipitating the supernatant with subsequent centrifuging to obtain a supernatant;

c) in the first or the second insulator

precipitating the supernatant;

centrifuging the precipitate;

solubilizing the pellet obtained by centrifuging the precipitate;

dialysing the solubilized pellet and centrifuging the dialysate;

performing chromatography of the dialysate;

filtration of the dialysate to obtain an eluate.