Purification of proteins

• US 10,233,211 B2
• Filed: 07/31/2013
• Issued: 03/19/2019
• Est. Priority Date: 12/21/2006
• Status: Active Grant

• Current Assignee: EMD Millipore Corporation (Merck & Co., Inc.)
• Original Assignee: EMD Millipore Corporation (Merck & Co., Inc.)
• Inventors: Moya, Wilson, Jaber, Jad
• Primary Examiner(s): Kolker, Daniel E
• Assistant Examiner(s): Rogers, James L
• Application Number:
  US13/955,024
• Publication Number:
  US 20130317204A1
• Time in Patent Office: 2,057 Days
• Field of Search: None
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 1/30 by precipitation
• CPC Class Codes: C07K 1/32 as complexes

Abstract

The present invention relates to a selectively soluble polymer capable of binding to a desired molecules in an unclarified mixture containing various biological materials and the methods of using such a polymer to purify a molecule from such a mixture. The polymer is soluble in the mixture under a certain set of process conditions such as pH or temperature and/or salt concentration and is rendered insoluble and precipitates out of solution upon a change in the process conditions. The polymer is capable of binding to the desired molecule (protein, polypeptide, etc) and remains capable of binding to that molecule even after the polymer is precipitated out of solution. The precipitate can then be filtered out from the remainder of the stream and the desired biomolecule is recovered such as by elution and further processed.

Claims

1. A method for purifying IgG antibody from an unclarified mixture containing impurities comprising:

a. providing the mixture at a set of conditions,

b. adding one or more polymers, soluble in said mixture under the set of conditions and capable of reversibly and selectively binding to at least the IgG antibody, said one or more polymers being one or more polyvinyl pyridine-containing polymers;

c. mixing the one or more solubilized polymers throughout the mixture;

d. precipitating the one or more polymers and at least the bound IgG antibody out of solution by changing the set of conditions in the mixture; and

e. separating the precipitated polymer and at least the bound IgG antibody from the mixture.

2. The method of claim 1 wherein the one or more polymers are precipitated by a change in pH.

3. The method of claim 1 wherein the one or more polymers are solubilized and precipitated by a change in pH.