Method for the reduction of host cell proteins in affinity chromatography
AssigneesGenentech, Inc., Hoffmann-La Roche Inc.
InventorsPaul McDonald, Richard St. John, Marc Wong, Roberto Falkenstein, Wolfgang Koehnlein, Klaus Schwendner, Bernhard Spensberger, Michael Wiedmann, Frank Zettl, Annika Kleinjans, Carina Kopp, Benjamin Tran, Ryan Erickson
Life of this patent
Estimating the term…
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.
Claims14
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.
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.
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.
Dependencies are inferred from the claim text.