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US 12,441,809 B2Granted
Affinity chromatography media for removal of anti-a and/or anti-b antibodies
AssigneeMerck Patent GmbH
InventorsNanying Bian, Chia-Yun Sun, Melissa Holstein, Kristen Cotoni, Matthew T. Stone, Santosh Rahane
Life of this patent
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Abstract
Embodiments described herein relate to novel chromatography media for removing anti-A and/or anti-B antibodies from a sample, as well as methods of using the same. The media described herein have several advantages over previously described media including, acid and alkaline stability.
Claims7
A method of removing anti-B antibodies from a sample, the method comprising the steps of:
- (a) providing a sample comprising an amount of anti-B antibodies;
- incubating the sample with a media for removing anti-B antibodies from a sample, the media comprising a solid support with a blood group B antigen oligosaccharide based ligand attached thereto, wherein the oligosaccharide based ligand is attached to the solid support at a ligand loading of at least 0.8 mg/ml of solid support, wherein the blood group B antigen ligand has the following structure:
- and wherein the media is stable under acid and alkaline conditions and wherein the solid support comprises a polyvinyl ether polymer and the blood group B antigen oligosaccharide based ligands are attached via pAA tentacle chemistry to a solid support, for the media to bind blood group B antibodies;
- (b) recovering portion of the sample which is not bound to the media;
- and
- (c) measuring amount of anti-B antibodies in the portion of the sample in (b), wherein the amount of anti-B antibodies in (c) is at least 80% less than the amount of anti-B antibodies in the sample in (a), wherein the media can be readily eluted and cleaned in place multiple times under a wide range of pH conditions, without losing its capability to remove the anti-B antibodies from the sample.
3 more elements
A method of purifying a monoclonal anti-B IgM antibody from a clarified cell culture feed, the method comprising the steps of:
- (a) providing a clarified cell culture feed containing a monoclonal anti-B IgM antibody;
- incubating the feed with a media for removing anti-B antibodies from a sample, the media comprising a solid support with a blood group B antigen oligosaccharide based ligand attached thereto, wherein the oligosaccharide based ligand is attached to the solid support at a ligand loading of at least 0.8 mg/ml of solid support, wherein the blood group B antigen ligand has the following structure:
- (b) and wherein the media is stable under acid and alkaline conditions and wherein the solid support comprises a polyvinyl ether polymer and the blood group B antigen oligosaccharide based ligands are attached via pAA tentacle chemistry to a solid support to facilitate the binding of anti-B IgM antibody to the media;
- (c) washing the media with an aqueous buffer having a pH ranging from 3.5 to 9.0, wherein the media can be readily eluted and cleaned in place multiple times without losing its capability to remove the anti-B IgM antibodies from the sample;
- (d) eluting the anti-B IgM antibody from the media using a buffer having a pH ranging from 2.0 to 3.0, thereby to obtain an eluate;
- and
- (e) recovering the purified anti-B IgM antibody in the eluate.
4 more elements
Dependencies are inferred from the claim text.