Stirrer system

Patent metadata

Field	Value
Publication number	US-2018187138-A1
Application number	US-201715800185-A
Country	US
Kind code	A1
Filing date	Nov 1, 2017
Priority date	Jul 24, 2009
Publication date	Jul 5, 2018
Grant date	—

Abstract

The present invention concerns a stirrer system for animal cell culture consisting of a combination of at least one radially-conveying stirrer element and at least one axially-conveying stirrer element, wherein at least three stirrer elements must be present and the uppermost stirrer element is an axially-conveying stirrer element. The stirrer elements are arranged at a certain distance above one another on a stirrer shaft. A particular embodiment is a multiple stirrer system consisting of two disk stirrers as radially-conveying stirrer elements and an inclined-blade stirrer as an axially-conveying stirrer element wherein the inclined-blade stirrer is arranged above the disk stirrer on the stirrer shaft. The stirrer system according to the invention achieves among others a gentler and better intermixing in the culture of shear-sensitive mammalian cells in cell cultures.

First claim

1 . A method for culturing mammalian cells, the method comprising culturing the mammalian cells in a device comprising a stirrer system and a cultivation vessel, wherein the stirrer system consists of two radially-conveying stirrer elements and one axially-conveying stirrer element arranged above one another on a vertical stirrer shaft, wherein the axially-conveying stirrer element is arranged above the two radially-conveying stirrer elements.

2 . The method of claim 1 , wherein the stirrer system has a Newton number of from 5.5 to 8.0 at a Reynolds number of from 5×10 4 to 5×10 5 when used in the cultivation vessel comprising cultivation medium.

3 . The method of claim 1 , wherein the stirrer system has a mixing time Θ 0.95 of about 20 seconds at a power input of about 0.05 W/kg and has a mixing time Θ 0.95 of about 10 seconds at a power input of about 0.3 W/kg when used in the cultivation vessel comprising cultivation medium.

4 . The method of claim 1 , wherein the stirrer system consists of two disk stirrers or two Rushton turbines as the radially-conveying stirrer elements and one inclined-blade stirrer as the axially-conveying stirrer element.

5 . The method of claim 1 , wherein the radially-conveying stirrer elements have between 2 and 8 stirrer blades and the axially-conveying stirrer element has between 2 and 10 stirrer blades.

6 . The method of claim 1 , wherein all the conveying stirrer elements have the same diameter d.

7 . The method of claim 1 , wherein the ratio of the diameter d of the stirring elements to the diameter D of the cultivation vessel is in the range of from 0.32 to 0.35.

8 . The method of claim 1 , wherein the pitch of the stirrer blades of the axially-conveying stirrer element is between 10° and 80° relative to the stirrer shaft.

9 . A method for producing a polypeptide, the method comprising culturing a mammalian cell comprising a nucleic acid encoding the polypeptide in the device of claim 1 , and recovering the polypeptide from the cultivation medium or from the cells, thereby producing the polypeptide.

10 . The method of claim 9 , wherein the stirrer system has a Newton number of from 5.5 to 8.0 at a Reynolds number of from 5×10 4 to 5×10 5 when used in the cultivation vessel comprising cultivation medium.

11 . The method of claim 9 , wherein the stirrer system has a mixing time Θ 0.95 of about 20 seconds at a power input of about 0.05 W/kg and has a mixing time Θ 0.95 of about 10 seconds at a power input of about 0.3 W/kg when used in the cultivation vessel comprising cultivation medium.

12 . The method of claim 9 , wherein the stirrer system consists of two disk stirrers or two Rushton turbines as the radially-conveying stirrer elements and one inclined-blade stirrer as the axially-conveying stirrer element.

13 . The method of claim 9 , wherein the radially-conveying stirrer elements have between 2 and 8 stirrer blades and the axially-conveying stirrer element has between 2 and 10 stirrer blades.

14 . The method of claim 9 , wherein all the conveying stirrer elements have the same diameter d.

15 . The method of claim 9 , wherein the ratio of the diameter d of the stirring elements to the diameter D of the cultivation vessel is in the range of from 0.32 to 0.35.

16 . The method of claim 9 , wherein the pitch of the stirrer blades of the axially-conveying stirrer element is between 10° and 80° relative to the stirrer shaft.

17 . A stirrer system consisting of two radially-conveying stirrer elements and one axially-conveying stirrer element arranged above one another on a vertical stirrer shaft, wherein the axially-conveying stirrer element is arranged above the radially-conveying stirrer elements.

18 . A device comprising the stirrer system of claim 17 and a cultivation vessel.

19 . The device of claim 18 , comprising a gas feed at the bottom of the cultivation vessel and at least one inlet for adding one or more solutions selected from the group consisting of a correcting solution and a feeding solution.

20 . The device of claim 18 or claim 19 , wherein the stirrer system has a Newton number of from 5.5 to 8.0 at a Reynolds number of from 5×10 4 to 5×10 5 when used in a cultivation vessel comprising cultivation medium.

21 . The device of claim 18 or claim 19 , wherein the stirrer system has a mixing time Θ 0.95 of about 20 seconds at a power input of about 0.05 W/kg and has a mixing time Θ 0.95 of about 10 seconds at a power input of about 0.3 W/kg when used in the cultivation vessel comprising cultivation medium.

22 . The device of claim 18 or claim 19 , wherein the stirrer system consists of two disk stirrers or two Rushton turbines as the radially-conveying stirrer elements and one inclined-blade stirrer as the axially-conveying stirrer element.

23 . The device of claim 18 or claim 19 , wherein the radially-conveying stirrer elements have between 2 and 8 stirrer blades and the axially-conveying stirrer element has between 2 and 10 stirrer blades.

24 . The device of claim 18 or claim 19 , wherein all the conveying stirrer elements have the same diameter d.

25 . The device of claim 18 or claim 19 , wherein the ratio of the diameter d of the stirring elements to the diameter D of the cultivation vessel is in the range of from 0.32 to 0.35.

26 . The device of claim 18 or claim 19 , wherein the pitch of the stirrer blades of the axially-conveying stirrer element is between 10° and 80° relative to the stirrer shaft.

Assignees

• Hoffmann La Roche

Inventors

• Alisch Alexander
• Jenzsch Marco
• Pohlscheidt Michael
• Thiele Joerg
• Wallerius Claus

Classifications

• C12N5/0602

  Vertebrate cells · CPC title

• C07K16/40

  against enzymes · CPC title

• C07K2317/14

  Specific host cells or culture conditions, e.g. components, pH or temperature · CPC title

• C12M27/08 Primary

  with different stirrer shapes in one shaft or axis · CPC title

• C07K16/2887

  against CD20 · CPC title