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AssigneeHoffmann-La Roche Inc.
InventorsRoberto Falkenstein, Maria Laura Magri, Michaela Mehr, Klaus Schwendner, Bernhard Spensberger
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Herein is reported a method for purifying a polypeptide comprising the steps of i) applying a solution comprising the polypeptide to an ion exchange chromatography material, and ii) recovering the polypeptide with a solution comprising a denaturant and thereby purifying the polypeptide, whereby the ion exchange chromatography material comprises a matrix of cross-linked poly (styrene-divinylbenzene) to which ionic ligands have been attached, and wherein the solution comprising the polypeptide applied to the ion exchange chromatography material is free of the denaturant and the polypeptide adsorbed to the ion exchange chromatography material is recovered with a solution comprising a denaturant at a constant conductivity.
) A method for purifying a polypeptide comprising the step of:
) A method for producing a polypeptide comprising the following steps:
) The method of claim 1, characterized in that the denaturant is one denaturant and is selected from the group comprising urea, guanidine, urea-derivatives, and guanidine-derivatives.
) The method of claim 2, characterized in that the denaturant is one denaturant and is selected from the group comprising urea, guanidine, urea-derivatives, and guanidine-derivatives.
) The method of claim 1, characterized in that the ion exchange chromatography material is a cation exchange chromatography material.
) The method of claim 2, characterized in that the ion exchange chromatography material is a cation exchange chromatography material.
) The method of claim 5, characterized in that the ionic ligand is a sulfopropyl ligand or a carboxymethyl ligand.
) The method of claim 6, characterized in that the ionic ligand is a sulfopropyl ligand or a carboxymethyl ligand.
) The method of claim 1, characterized in that the ion exchange chromatography material is an anion exchange chromatography material.
) The method of claim 2, characterized in that the ion exchange chromatography material is an anion exchange chromatography material.
) The method of claim 9, characterized in that the ionic ligand is an ethyleneimine ligand or a quaternized ligand.
) The method of claim 10, characterized in that the ionic ligand is an ethyleneimine ligand or a quaternized ligand.
) The method of claim 1, further characterized in comprising the following steps:
) The method of claim 2, further characterized in comprising the following steps:
) The method according to claim 13, characterized in that
) The method according to claim 14, characterized in that
) The method of claim 13, characterized in that the recovered polypeptide is refolded prior to the applying to the second ion exchange chromatography material.
) The method of claim 14, characterized in that the recovered polypeptide is refolded prior to the applying to the second ion exchange chromatography material.
) The method of claim 1, wherein the purifying further comprises the following steps:
The method of claim 2, wherein the purifying further comprises the following steps:
) The method according to claim 19, characterized in that after applying the second solution and prior to applying the fourth solution the following step is added:
) The method of claim 1, characterized in that the polypeptide has an amino acid sequence selected from the group of amino acid sequences of SEQ ID NO:
) The method of claim 2, characterized in that the polypeptide has an amino acid sequence selected from the group of amino acid sequences of SEQ ID NO:
) The method of claim 1, wherein the denaturant is a urea or a urea-derivate.
) The method of claim 2, wherein the denaturant is a urea or a urea-derivate.
Dependencies are inferred from the claim text.