Stimulus responsive polymers for the purification of biomolecules

Patent metadata

Field	Value
Publication number	US-9731288-B2
Application number	US-201514940864-A
Country	US
Kind code	B2
Filing date	Nov 13, 2015
Priority date	May 17, 2010
Publication date	Aug 15, 2017
Grant date	Aug 15, 2017

Abstract

The present invention provides novel and improved stimulus responsive polymers and methods of using the same for the purification of biomolecules.

First claim

What is claimed is:

1. A method of separating a target molecule from one or more impurities in a sample, where the method comprises the steps of:
(a) providing a sample comprising a target molecule and one or more impurities;
(b) contacting the sample with a soluble stimulus responsive polymer selected from the group consisting of benzyl modified polyallylamine; diphenyl modified polyallylamine; dichlorobenzyl modified polyallylamine; chlorobenzyl modified polyallylamine; hexanoic acid and tert-butyl modified polyallylamine; phenylbenzyl modified polyallylamine; benzyl modified polyethyleneimine; and benzyl modified polyvinylamine, wherein the stimulus responsive polymer is capable of binding and precipitating one or more impurities following the addition of a multivalent ion stimulus; and
(c) adding a multivalent ion stimulus selected form phosphate or citrate to the sample to precipitate complex of polymer and one or more impurities out of the sample;
thereby to separate the target molecule from one or more impurities.

2. The method of claim 1 , wherein the target molecule is a recombinant protein.

3. The method of claim 1 , wherein the target molecule is an antibody.

4. The method of claim 3 , wherein the antibody is a monoclonal antibody.

5. The method of claim 1 , wherein the one or more impurities is selected from the group consisting of host cell protein, DNA, RNA, viruses, antibody aggregates and cell culture additives.

6. The method of claim 1 , wherein the sample comprises a feedstock.

7. The method of claim 1 , wherein the sample comprises cell culture media into which the target molecule is secreted.

8. The method of claim 7 , wherein the cell culture media is a CHO cell culture media.

9. The method of claim 7 , wherein the target molecule is a monoclonal antibody.

10. The method of claim 1 , further comprising one or more chromatography steps.

11. The method of claim 1 , further comprising one or more filtration steps.

12. The method of claim 1 , further comprising the step of removing a residual amount of polymer after step (c), wherein the step comprises the use of a membrane or resin modified with a multivalent ion.

13. The method of claim 12 , wherein the multivalent ion is phosphate.

Assignees

• Emd Millipore Corp

Inventors

• Jaber Jad
• Moya Wilson
• Hamzik James
• Boudif Arezki
• Zhang Yu
• Soice Neil
• Charkoudian John
• Singh Nripen

Classifications

• B01J41/13

  obtained otherwise than by reactions only involving unsaturated carbon-to-carbon bonds · CPC title

• C08F8/00

  Chemical modification by after-treatment (graft polymers, block polymers, crosslinking with unsaturated monomers or with polymers C08F251/00 - C08F299/00; of conjugated diene rubbers C08C) · CPC title

• C07K1/30

  by precipitation · CPC title

• C07K2317/10

  characterized by their source of isolation or production · CPC title

• B01J41/14 Primary

  obtained by reactions only involving unsaturated carbon-to-carbon bonds · CPC title