Removal of microorganisms from cell culture media

Patent metadata

Field	Value
Publication number	US-2016177252-A1
Application number	US-201514870519-A
Country	US
Kind code	A1
Filing date	Sep 30, 2015
Priority date	Dec 22, 2014
Publication date	Jun 23, 2016
Grant date	—

Abstract

Compositions and methods are provided for removing viral contaminants from a chemically defined cell culture medium. Compositions provided herein are resistant to or exhibit reduced fouling by one or more components in a chemically defined cell culture medium.

First claim

What is claimed is:

1 . A porous membrane having a surface modified with a polymer comprising randomly arranged and crosslinked monomers of diacetone acrylamide and one or more non-acrylamide cross-linkable monomers.

2 . The membrane of claim 1 , wherein the polymer is directly coated on the surface of the porous membrane using an energy source.

3 . The membrane of claim 1 , wherein the energy source is selected from the group consisting of heat, electron beam, ultraviolet light and gamma radiation.

4 . The membrane of claim 1 , wherein a non-acrylamide cross-linkable monomer is polyethylene glycol diacrylate

5 . The porous membrane of claim 1 , wherein the porous membrane is an asymmetric membrane.

6 . The porous membrane of claim 5 , wherein the asymmetric membrane is a polyethersulfone (PES) membrane.

7 . A method of removing a virus contaminant from a chemically defined cell culture medium comprising filtering the chemically defined cell culture medium through the porous membrane of claim 1 .

8 . A porous asymmetric PES membrane modified with a polymer comprising the following structure:

wherein,
M1 is neutral DACm monomer: H 2 C═CH—C(O)—NH—C(CH 3 ) 2 —CH 2 —C(O)—CH 3 ;
M2 is neutral PEGDA monomer: H 2 C═CH—C(O)—O—(CH 2 —CH 2 —O) n —C(O)—CH═CH 2 ;
M1* is radicalized DACm monomer: *H 2 C—CH—C(O)—NH—C(CH 3 ) 2 —CH 2 —C(O)—CH 3 ;
M2′ is the radicalized PEGDA monomer: *H 2 C—CH—C(O)—O—(CH 2 —CH 2 —O) n —C(O)—CH—CH 2 * ;
P refers to a random polymer network;
n=6, 7, 8, 9, 10, 11; and
the symbol “*” refers to an alkenyl radical.

9 . A porous asymmetric PES membrane modified with a polymer comprising the following structure:

wherein,
M1 and M2 refer to a neutral PEGDA monomer: H 2 C═CH—C(O)—O—(CH 2 —CH 2 —O) n —C(O)—CH═CH 2 ;
Mr and M2′ refer to a radicalized PEGDA monomer: *H 2 C—CH—C(O)—O—(CH 2 —CH 2 —O) n —C(O)—CH—CH 2 * ;
P refers to a random polymer network;
n=6, 7, 8, 9, 10, 11; and
the symbol “*” refers to an alkenyl radical.

10 . A method of removing one or more viral contaminants from a chemically defined cell culture medium, the method comprising the steps of:
a) providing a chemically defined cell culture medium; and b) filtering the chemically defined cell culture medium through the porous membrane of claim 1 , prior to or during transfer of the medium into a bioreactor; wherein the level of one or more viral contaminants in the chemically defined cell culture medium inside the bioreactor is lower than the level prior to filtering the medium through the membrane.

11 . The method of claim 10 , wherein the membrane is incorporated into a device.

12 . The method of claim 11 , wherein the device is in a format selected from a disc, a pleated cartridge, a spirally wound cartridge and a multi-plate flat sheet.

13 . The method of claim 7 , wherein the chemically defined cell culture medium is selected from the group consisting of Lonza Power CHO, CD Opti CHO, EMD Millipore Cellvento CHO 100 and Cellvento CHO 200.

14 . The method of claim 10 , wherein the chemically defined cell culture medium is selected from the group consisting of Lonza Power CHO, CD Opti CHO, EMD Millipore Cellvento CHO 100 and Cellvento CHO 200.

15 . The method of claim 10 , wherein level of one or more viral contaminants following step (b) is reduced by at least 1 Logio reduction value (LRV) or at least 4 Logio reduction value (LRV) or at least 6 Logio reduction value (LRV).

16 . The method of claim 7 , wherein the filtration is carried out for a period of less than 24 hours.

17 . The method of claim 7 , wherein the filtration is carried out for a period of less than 24 hours.

18 . The method of claim 10 , wherein the filtration is carried out for a period of less than 24 hours.

19 . The method of claim 7 , wherein filtration is conducted at a pH ranging from 4 to 8.

20 . The method of claim 10 , wherein filtration is conducted at a pH ranging from 4 to 8.

21 . The method of claim 7 , wherein filtration is conducted at a temperature ranging from 20° C. to 25° C.

22 . The method of claim 10 , wherein normal flow filtration is conducted at a temperature ranging from 20° C. to 25° C.

23 . A method of reducing the fouling of a virus retentive membrane by one or more components in a chemically defined cell culture medium during filtration, the method comprising the steps of:
a) providing a virus retentive membrane; and b) modifying the membrane with a polymer comprising randomly arranged and crosslinked monomers of diacetone acrylamide and one or more cross-linkable monomers, wherein the fouling of the modified membrane by one or more components in a chemically defined cell culture medium is reduced relative to an unmodified membrane.

24 . The method of claim 23 , wherein the virus retentive membrane is a PES membrane, a PVDF membrane, a cellulosic membrane or a nylon membrane.

25 . The method of claim 23 , wherein a non-acrylamide cross-linkable monomer is PEGDA.

Assignees

• Emd Millipore Corp

Inventors

• Peters Antoni
• Goddard Philip M

Classifications

• A61L2/022 Primary

  Filtration · CPC title

• B01D67/0088

  Physical treatment with compounds, e.g. swelling, coating or impregnation · CPC title

• B01D67/009

  with wave-energy, particle-radiation or plasma · CPC title

• B01D71/82

  characterised by the presence of specified groups, e.g. introduced by chemical after-treatment · CPC title

• B01D61/14

  Ultrafiltration; Microfiltration · CPC title