Optimized method for antibody capturing by mixed mode chromatography
US-9422329-B2 · Aug 23, 2016 · US
Method for separation of monomeric polypeptides from aggregated polypeptides
US9783570B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9783570-B2 |
| Application number | US-201214127085-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jun 28, 2012 |
| Priority date | Jul 1, 2011 |
| Publication date | Oct 10, 2017 |
| Grant date | Oct 10, 2017 |
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Abstract
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Herein is reported a method for obtaining a polypeptide in monomeric form comprising the steps of a) providing a solution comprising the polypeptide m monomeric form and m aggregated form, wherein the ratio of monomeric to aggregated form is 4:1 or less as determined by size exclusion chromatography, b) performing a mixed-mode chromatography in bind-and-elute mode, or a hydrophobic interaction chromatography in flow-through mode, or a size-exclusion chromatography, and c) performing a weak cation exchange chromatography in bind-and-elute mode or flow-through mode, and thereby obtaining the polypeptide m monomeric form.
First claim
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The invention claimed is: 1. A method for purifying a monomeric Fc-region containing polypeptide comprising the steps: a) applying a solution comprising a Fc-region containing polypeptide in monomeric form and in aggregated form to a mixed-mode chromatography material, wherein the aggregated form is 20% or more as determined by size exclusion chromatography of the total Fc-region containing polypeptide in the solution, wherein said solution has a pH value of about 0.2 to about 2.5 pH units above the pl of the polypeptide, and recovering a solution comprising about 95% monomeric Fc-region containing polypeptide; and b) applying the solution recovered in step a) to a weak cation exchange chromatography material, wherein said solution has pH value of about 0.2 to about 2.5 pH units above the pl of the polypeptide, and recovering the monomeric form of the Fc-region containing polypeptide. 2. The method according to claim 1 , wherein the monomeric form of the Fc-region containing polypeptide is recovered from the flow-through of the weak cation exchange chromatography material of step b). 3. The method according to claim 1 , wherein the solution applied to the mixed-mode chromatography material of step a) and/or the weak cation exchange chromatography material of step b) is buffered with a citrate buffer, acetate buffer, and/or MES buffer.
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Classifications
- C07K1/165Primary
mixed-mode chromatography · CPC title
by a combination of two or more processes of different types · CPC title
Ion-exchange chromatography · CPC title
Partition-, reverse-phase or hydrophobic interaction chromatography · CPC title
IL-15 · CPC title
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- What does patent US9783570B2 cover?
- Herein is reported a method for obtaining a polypeptide in monomeric form comprising the steps of a) providing a solution comprising the polypeptide m monomeric form and m aggregated form, wherein the ratio of monomeric to aggregated form is 4:1 or less as determined by size exclusion chromatography, b) performing a mixed-mode chromatography in bind-and-elute mode, or a hydrophobic interaction …
- Who is the assignee on this patent?
- Baehner Monika, Grossmann Adelbert, Hepbildikler Stefan, and 1 more
- What technology area does this patent fall under?
- Primary CPC classification C07K1/165. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Oct 10 2017 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 2 related publications on this page (citations in our corpus or others sharing the same primary CPC).