Method for the reduction of host cell proteins in affinity chromatography

Patent metadata

Field	Value
Publication number	US-12503487-B2
Application number	US-202117328408-A
Country	US
Kind code	B2
Filing date	May 24, 2021
Priority date	Aug 21, 2015
Publication date	Dec 23, 2025
Grant date	Dec 23, 2025

Abstract

The current invention reports a method for purifying an antibody by reducing the content of a host cell protein. The method employs a wash step with a low conductivity aqueous solution in an affinity chromatography.

First claim

The invention claimed is:

1 . A method of reducing content of a host cell protein in a sample including a human IgG4 or IgG1 isotype antibody and the host cell protein, the method comprising the steps of:
contacting a protein A chromatography material with the sample; washing the protein A chromatography material, after the contacting step, with a low conductivity aqueous solution, wherein the low conductivity aqueous solution has a conductivity value of about 0.5 mS/cm or less, wherein the amount of the host cell protein in the sample after the washing step is reduced, wherein said host cell protein is phospholipase B-like 2 (PLBL2), and wherein the low conductivity aqueous solution is deionized water.

2 . The method according to claim 1 , wherein the low conductivity aqueous solution wash step is preceded or succeeded by a high conductivity aqueous solution wash step.

3 . The method according to claim 2 , wherein the high conductivity aqueous solution has a conductivity value of about 20 mS/cm or higher.

4 . The method according to claim 2 or 3 , wherein the high conductivity aqueous solution comprises Histidine.

5 . The method according to claim 1 , wherein the human IgG4 isotype antibody is an antibody against P-selectin or an antibody against factor IXa and factor X or an antibody against IL-13 or an antibody against amyloid beta.

6 . The method according to claim 1 , wherein the human IgG1 isotype antibody is an antibody against Influenza B or an antibody against VEGF-A or an antibody against CD22 or a bispecific antibody against HER3 and EGFR or an antibody against amyloid beta or an antibody against Her2 or a bispecific antibody against Ang2 and VEGF-A or a bispecific antibody against carcinoembryonic antigen (CEA) and CD3.

7 . A method for producing a human IgG4 or IgG1 isotype antibody comprising the steps of:
a) cultivating a cell comprising a nucleic acid encoding a human IgG4 or IgG1 isotype antibody, b) recovering the human IgG4 or IgG1 isotype antibody from the cell or the cultivation medium, c) contacting the human IgG4 or IgG1 isotype antibody with a protein A chromatography material, d) washing the protein A chromatography material, after the contacting step, with a low conductivity aqueous solution, wherein the low conductivity aqueous solution has a conductivity value of about 0.5 mS/cm or less, wherein the amount of a host cell protein in the sample after the washing step is reduced, wherein said host cell protein is phospholipase B-like 2 (PLBL2), e) recovering the human IgG4 or IgG1 isotype antibody from the protein A chromatography material and thereby producing the human IgG4 or IgG1 isotype antibody; wherein the low conductivity aqueous solution is deionized water.

8 . A method for purifying a human IgG4 or IgG1 isotype antibody from a sample comprising the steps of:
a) providing a sample comprising a human IgG4 or IgG1 isotype antibody, b) purifying the human IgG4 or IgG1 isotype antibody with a protein A chromatography method/step, comprising contacting the sample with a protein A chromatography material and washing the protein A chromatography material, after the contacting step, with a low conductivity aqueous solution, wherein the low conductivity aqueous solution has a conductivity value of about 0.5 mS/cm or less, wherein the amount of a host cell protein in the sample after the washing step is reduced and wherein said host cell protein is phospholipase B-like 2 (PLBL2), wherein the low conductivity aqueous solution is deionized water.

9 . The method according to claims 7 or 8 , wherein the method additionally comprises washing the affinity chromatography material with a high conductivity aqueous solution and/or with a medium conductivity aqueous solution before or after washing the protein A chromatography material with low conductivity aqueous solution.

10 . The method according to claim 9 , wherein the high conductivity aqueous solution has a conductivity value of about 20 mS/cm or higher.

11 . The method according to claim 9 , wherein the medium conductivity aqueous solution has a conductivity value of from more than 0.5 mS/cm to less than 20 mS/cm.

12 . The method according to claim 9 , wherein the high or medium conductivity aqueous solution comprises Histidine.

13 . The method according to claims 7 or 8 , wherein the human IgG4 isotype antibody is an antibody against P-selectin or an antibody against factor IXa and factor X or an antibody against IL-13 or an antibody against amyloid beta.

14 . The method according to claims 7 or 8 , wherein the human IgG1 isotype antibody is an antibody against Influenza B or an antibody against VEGF-A or an antibody against CD22 or a bispecific antibody against HER3 and EGFR or an antibody against amyloid beta or an antibody against Her2 or a bi specific antibody against Ang2 and VEGF-A or a bispecific antibody against carcinoembryonic antigen (CEA) and CD3.

Assignees

• Genentech Inc
• Hoffmann La Roche

Inventors

• Mcdonald Paul
• St John Richard
• Wong Marc
• Falkenstein Roberto
• Koehnlein Wolfgang
• Schwendner Klaus
• Spensberger Bernhard
• Wiedmann Michael
• Zettl Frank
• Kleinjans Annika
• Kopp Carina
• Tran Benjamin
• Erickson Ryan

Classifications

• C07K16/108

  Orthomyxoviridae (F), e.g. influenza virus · CPC title

• C07K2317/14

  Specific host cells or culture conditions, e.g. components, pH or temperature · CPC title

• C07K2317/31

  multispecific · CPC title

• C07K16/36

  against blood coagulation factors · CPC title

• C07K16/32

  against translation products of oncogenes · CPC title