Granted Patent B2

US 12,441,809 · App. 17/088,659 · Granted Oct 14, 2025

Inventors: Nanying Bian (Lexington, MA); Chia-Yun Sun (Arlington, MA); Melissa Holstein (Concord, MA); Kristen Cotoni (Malden, MA); Matthew T. Stone (Arlington, MA); Santosh Rahane (Acton, MA)

Assignee: Merck Patent GmbH

C07K16/34B01D15/20B01D15/3809B01D15/426B01J20/285B01J20/289B01J20/321B01J20/3274C07K1/22B01J2220/54B01J2220/58C07K2317/21C07K2317/70

Abstract

Claims (17)

1. 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.

2. The method according to claim 1 , wherein the ligand loading of the media is at least 1 mg/ml of solid support or at least 1.2 mg/ml of solid support.

3. The method of claim 1 , wherein the media is packed in a device.

4. The method of claim 3 , wherein the device is a chromatography column.

5. 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.

6. The method of claim 5 , wherein the media are packed in a chromatography column.

7. The method of claim 6 , wherein the clarified cell culture feed is flowed through the column.

Continuity (3)

References Cited (108)