Cell retention device and method

Patent metadata

Field	Value
Publication number	US-2019169559-A1
Application number	US-201716092482-A
Country	US
Kind code	A1
Filing date	Apr 13, 2017
Priority date	Apr 15, 2016
Publication date	Jun 6, 2019
Grant date	—

Abstract

The subject technology relates to a cell retention device and method for use in a perfusion cell culture system where the cell retention device includes a hollow-fiber filter having an average pore size ranging from about 0.5 to about 20 μm and having the ability to operate under perfusion cell culture conditions for up to 35 days without being clogged or losing its product sieving ability by more than 20%.

First claim

1 . A cell retention device comprising a hollow-fiber filter having an average pore size ranging from about 0.5 to about 20 μm.

2 . The cell retention device of claim 1 , wherein the cell retention device is external to a perfusion cell culture vessel.

3 . The cell retention device of claim 1 , wherein the hollow-fiber filter has an average pore size selected from about 5 to about 8 μm, 5 μm to 15 μm, 5.2 μm to 12 μm, 5.5 μm to 8 μm, 5.2 μm to 7.7 μm, 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 11 μm, 12 μm, 13 μm, 15 μm, or any size above 5 μm that could retain the viable cells while allowing dead cells, cellular debris or macromolecules to pass through the filter.

4 . The cell retention device of claim 1 , wherein the hollow-fiber filter has an average pore size that is equal or greater than 5 μm but less than the size of viable cells being cultured in a cell culture vessel.

5 . The cell retention device of claim 1 , wherein the hollow-fiber filter has an average pore size greater than the size of a viable cell by 5 μm or less.

6 . The cell retention device of claim 1 , wherein the cell retention device is configured to operate by tangential-flow filtration (TFF) or by alternating tangential flow filtration (ATF).

7 . The cell retention device of claim 5 , wherein the TTF or ATF are configured to operate under a low shear rate of about 1000 s −1 to about 4000 s −1 .

8 . The cell retention device of claim 1 , wherein the hollow-fiber filter has filter capacity of about 1000 to about 10,000 L/M 2 .

9 . The cell retention device of claim 1 , wherein the cell retention device is configured to operate in conjunction with a perfusion culture vessel.

10 . The cell retention device of claim 1 , wherein the hollow-fiber filter is made out of ceramic, polymer or metallic materials.

11 . The cell retention device of claim 1 , wherein the hollow-fiber filter operates for up to 35 days without being clogged or losing more than 10% of its product sieving ability.

12 . The cell retention device of claim 10 , wherein the hollow-fiber filter operates in TTF or ATF modes under a low shear rate of 2000 s −1 or less or a perfusion flow rate of equal or greater than 4000 Um 2 .

13 . The cell retention device of claim 10 , wherein the hollow-fiber filter is in a housing that facilitates the collection of filtrates.

14 . A method for harvesting recombinant protein products from a perfusion culture vessel, said method comprising the steps of:
a. subjecting cell culture fluid of a perfusion culture vessel to a cell retention device to be filtered, wherein the cell culture fluid comprises cells and recombinant protein products produced by said cells and wherein the cell retention device comprises a hollow-fiber filter having an average pore size ranging from about 0.5 to about 20 μm; b. collecting filtrate from the cell retention device, wherein the filtrate comprises the recombinant protein products; c. Recirculating the filtered cell culture fluid to the perfusion culture vessel.

15 . The method of claim 14 , wherein the cell retention device is external to the perfusion cell culture vessel.

16 . The method of claim 14 , wherein the hollow-fiber filter filters the cell culture fluid by tangential-flow filtration (TFF) or alternating tangential flow filtration (ATF).

17 . The method of claim 14 , wherein the hollow-fiber filter has an average pore size selected from about 5 to about 8 micron, 5 μm to 15 μm, 5.2 μm to 12 μm, 5.5 μm to 8 μm, 5.2 μm to 7.7 μm, 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 11 μm, 12 μm, 13 μm, 15 μm, or any size above 5 μm that could retain the viable cells while allowing dead cells, cellular debris or macromolecules to pass through the filter.

18 . The method of claim 14 , wherein the hollow-fiber filter has an average pore size that is equal or greater than 5 micron but less than the size of the cells.

19 . The method of claim 14 , wherein the hollow-fiber filter has an average pore size greater than the size of the cells by 5 μm or less.

20 . The method of claim 14 , where in the hollow-fiber filter has an average pore size that is about the size of the cells.

21 - 46 . (canceled)

Assignees

• Boehringer Ingelheim Int

Inventors

• Coffman jonathan
• Godfrey Scott
• Wang Samantha

Classifications

• C12M29/16

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

• C12M29/10 Primary

  Perfusion · CPC title

• C12M47/02

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

• C12M47/10

  Separation or concentration of fermentation products (bioreactors combined with means for distillation or extraction of liquid fuel C12M43/02) · CPC title