# US 8,772,018 B2 · Media for affinity chromatography

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- Number: US 8,772,018 B2
- Document ID: US8772018B2
- Status: Granted
- Kind: B2
- Application number: 12/217,558
- Filed: 2008-07-07
- Published: 2009-10-01
- Granted: 2014-07-08
- Earliest claimed: 2007-07-10
- Assignees: EMD Millipore Corporation
- Applicants: Nanying Bian; Senthil Ramaswamy; Neil Soice; Chen Wang; Yuan Wong
- Inventors: Nanying Bian; Senthil Ramaswamy; Neil Soice; Chen Wang; Yuan Wong
- Web page: https://patentgrep.com/patent/US8772018B2

## CPC

- C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
- B01J 20/103: {comprising silica}
- B01J 20/28004: {Sorbent size or size distribution, e.g. particle size}
- B01J 20/28014: {characterised by their form}
- B01J 20/28016: {Particle form}
- B01J 20/28026: {Particles within, immobilised, dispersed, entrapped in or on a matrix, e.g. a resin}
- B01J 20/28054: {characterised by their surface properties or porosity}
- B01J 20/28085: {being more than 50 nm, i.e. macropores}
- B01J 20/281: Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J 20/283: based on silica
- B01J 20/286: Phases chemically bonded to a substrate, e.g. to silica or to polymers
- C03C 11/00: Multi-cellular glass {; Porous or hollow glass or glass particles}
- C03C 12/00: Powdered glass (C03C8/02 takes precedence); Bead compositions
- C03C 17/28: with organic material (C03C17/34, C03C17/44 take precedence)
- C07K 1/22: Affinity chromatography or related techniques based upon selective absorption processes
- G01N 33/538: by sorbent column, particles or resin strip {, i.e. sorbent materials}
- Y10T 428/2982: Particulate matter [e.g., sphere, flake, etc.]
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- C03C: CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

The invention relates generally to solid supports for chromatography. In specific embodiments the invention provides for solid supports suitable for affinity chromatography along with methods, systems and kits which use the same.

## Claims

### Claim 1

A solid support suitable for performing affinity chromatography comprising
i) a silica particle having a pore size greater than 630 Å and less than 1000 Å and a mean silica particle size greater than 50 μm and less than 70 μm, and
ii) an affinity ligand linked to the solid support wherein the affinity ligand is selected from the group consisting of proteins, enzymes, antibodies, immunoglobulins, and protein receptors and the affinity ligand density is 12 g/L to 14 g/L.

### Claim 2 (depends on 1)

The solid support of claim 1, wherein the silica particle is a bead.

### Claim 3 (depends on 1)

The solid support of claim 1, wherein the silica particle has a pore size ranging from 782 Å to 932 Å.

### Claim 4 (depends on 1)

The solid support of claim 1, wherein the silica particle has a mean particle size ranging from 55 μm to 67 μm.

### Claim 5 (depends on 1)

The solid support of claim 1, wherein the silica particle is comprised of a material chosen from particulate glass, controlled porous glass, colloidal silica, wollastonite, dried silica gel and bentonite.

### Claim 6 (depends on 1)

The solid support of claim 1, wherein the affinity ligand density is 13 g/L.

### Claim 7 (depends on 1)

The solid support of claim 1, wherein the dynamic capacity of the affinity ligand for a target molecule is 50 g/L to 60 g/L.

### Claim 8 (depends on 1)

The solid support of claim 1, wherein the affinity ligand specifically binds to a portion of an immunoglobulin.

### Claim 9 (depends on 8)

The solid support of claim 8, wherein the affinity ligand binds to a portion of an Fc region of an immunoglobulin.

### Claim 10 (depends on 8)

The solid support of claim 8, wherein the affinity ligand is chosen from Protein A, Protein G or a functional variant of either.

### Claim 11

A kit comprising an affinity chromatography solid support comprising a silica particle having a pore size greater than 630 Å and less than 1000 Å and a mean particle size greater than 55 μm and less than 70 μm, an affinity ligand linked to the solid support having an affinity ligand density from greater than 15 g/L, and a dynamic capacity for a target molecule 50 g/L to 60 g/L, wherein the affinity ligand is selected from the group consisting of proteins, enzymes, antibodies, immunoglobulins, and protein receptors, and
and at least one container.

### Claim 12 (depends on 11)

The kit of claim 11, further comprising a column suitable for packing the affinity chromatography solid support.

### Claim 13

A solid support suitable for performing affinity chromatography comprising
i) a silica bead selected from the group consisting of a particulate glass, controlled porous glass, colloidal silica, wollastonite, dried silica gel and bentonite, having a pore size from about 782 Å to 932 Å and a mean particle size from about 55 μm to 67 μm, and
ii) an affinity ligand, having an affinity ligand density from 12 g/L to 14 g/L, linked to the solid support wherein the affinity ligand is selected from the group consisting of proteins, enzymes, antibodies, immunoglobulins, and protein receptors,
wherein the affinity ligand has a dynamic capacity for a target molecule at least 50 g/L.

### Claim 14

A kit comprising an affinity chromatography solid support and a column suitable for packing the affinity chromatography solid support,
the affinity chromatography solid support comprises a silica particle having a pore size greater than about 782 Å to 932 Å and a mean particle size from 55 μm to 67 μm, an affinity ligand linked to the solid support having an affinity ligand density from 12 g/L to 14 g/L, and a dynamic capacity for a target molecule 50 g/L to 60 g/L, wherein the affinity ligand is selected from the group consisting of proteins, enzymes, antibodies immunoglobulins, and protein receptors, and at least one container.