Purification of proteins

• US 9,376,464 B2
• Filed: 01/23/2013
• Issued: 06/28/2016
• 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
• Application Number:
  US13/747,495
• Publication Number:
  US 20130137860A1
• Time in Patent Office: 1,252 Days
• Field of Search: None
• US Class Current: 1/1
• 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 biomolecules in a mixture containing various biological materials and the methods of using such a polymer to purify a biomolecule 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 biomolecule (protein, polypeptide, etc) and remains capable of binding to that biomolecule 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 an IgG antibody produced in a microbial host cell, the method comprising the steps of:

a. providing a mixture comprising an IgG antibody produced in a microbial host cell;

b. adding one or more stimuli responsive polymers to the mixture under pH conditions such that the one or more polymers are solubilized and are capable of reversibly and selectively binding to the IgG antibody, said one or more polymers being one or more polyvinyl pyridine-containing polymers;

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

d. changing the pH conditions such that to precipitate the one or more polymers and bound IgG antibody out of solution; and

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

2. The method of claim 1, further comprising the step of recovering the bound IgG antibody from the polymer.

3. The method of claim 1, wherein the microbial host cell is a yeast cell.

4. The method of claim 1, wherein the microbial host cell is an E. coli cell.