Method for membrane gas transfer in high density bioreactor culture

Patent metadata

Field	Value
Publication number	US-12415980-B2
Application number	US-201917283273-A
Country	US
Kind code	B2
Filing date	Oct 8, 2019
Priority date	Oct 10, 2018
Publication date	Sep 16, 2025
Grant date	Sep 16, 2025

Abstract

The present invention provides improved bioprocessing systems and methods for cell culture using the improved bioreactors, e.g., batch-fed or perfusion bioreactor cell culture systems for production of monoclonal or bi-specific antibodies, which are modified to include one or more membrane gas transfer modules in place of a sparger- or microsparger-based aeration systems to better regulate the levels of critical gases in a bioreactor cell culture, e.g., the dissolved levels of O 2 and CO 2 , even at high cell densities, without subjecting the cells to bubble-burst associated cell death.

First claim

The invention claimed is:

1. A method of culturing Chinese Hamster Ovary (CHO) cells in a bioreactor comprising providing a mass transfer of a gas to/from the bioreactor without generating bubbles inside the bioreactor, wherein the mass transfer of gas to/from the bioreactor is provided by two or more gas transfer modules comprising a plurality of hollow fibers, wherein at least one gas transfer module comprises an oxygen flow path through the hollow fibers and a cell culture medium flow path around the hollow fibers separated by a non-porous membrane comprising PDMS and wherein at least one gas transfer module comprises an air/carbon dioxide flow path through the hollow fibers and a cell culture medium flow path around the hollow fibers separated by a non-porous membrane comprising PDMS; wherein the bioreactor is a perfusion bioreactor, and wherein the bioreactor comprises a CHO cell density of at least 20×10 6 cells/ml.

2. The method of claim 1 , wherein the two or more gas transfer modules are located outside of the bioreactor.

3. The method of claim 2 , wherein the flow of cell culture media and/or cells comprises tangential, axial flow or a combination thereof.

4. The method of claim 2 , wherein the flow of the cell culture media and/or cells is at a rate that is sufficient to maintain culture homogeneity without causing shear forces on the cells.

5. The method of claim 1 , wherein the plurality of hollow fibers provide a flow path for culture media and cells to travel through spaces separating the hollow fibers.

6. The method of claim 5 , wherein the spaces are homogenous or heterogenous.

7. The method of claim 5 , wherein the spaces are of sufficient size to allow passage of a cell without causing shear forces on the cell.

8. The method of claim 5 , wherein the spaces comprise a distance of about 15 μm to about 2000 μm.

9. The method of claim 8 , wherein the spaces comprise a distance 15-30 μm, 20-40 μm, 30-60 μm, 40-80 μm, 60-120 μm, 80-160 μm, 100-200 μm, 150-300 μm, 200-400 μm, 200-500 μm, 200-600 μm, 200-700 μm, 200-800 μm, 200-900 μm, 200-1000 μm, or 500-2000 μm, or a combination thereof.

10. The method of claim 1 , wherein the bioreactor comprises a cell density of about 30×10 6 cells/ml.

11. The method of claim 1 , wherein the method avoids production of foam.

12. The method of claim 1 , wherein the bioreactor comprises no headspace or substantially no headspace.

13. The method of claim 1 , wherein the method requires no anti-foaming agent during cell culture.

14. The method of claim 1 , wherein a dissolved oxygen content in the bioreactor is maintained at around 60%.

15. The method of claim 14 , wherein hollow fibers have a wall thickness of 55 μm.

Assignees

• Boehringer Ingelheim Int

Inventors

• Coffman jonathan
• Godfrey Scott
• Sawicki Lisa

Classifications

• C12M47/02

  Separating microorganisms from the culture medium; Concentration of biomass (separating microorganisms from their culture media C12N1/02) · CPC title

• C12M41/34

  of gas · CPC title

• C12M41/26

  of pH · CPC title

• C12M41/02

  of foam (foam prevention during gasification of liquids B01D19/02) · CPC title

• C12M29/16

  Hollow fibers (hollow fiber modules in general B01D63/02) · CPC title