# US 8,465,935 B2 · Detection and quantitation of cyclodextrins

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 8,465,935 B2
- Document ID: US8465935B2
- Status: Granted
- Kind: B2
- Application number: 13/536,069
- Filed: 2012-06-28
- Published: 2013-01-03
- Granted: 2013-06-18
- Earliest claimed: 2004-12-06
- Assignees: Biogen Idec MA Inc.
- Applicants: Zoran Sosic; Rulin Qian; James Ahern; Rohin Mhatre
- Inventors: Zoran Sosic; Rulin Qian; James Ahern; Rohin Mhatre
- Web page: https://patentgrep.com/patent/US8465935B2

## CPC

- C08B 37/0015: {Inclusion compounds, i.e. host-guest compounds, e.g. polyrotaxanes}
- G01N 2030/8813: {biological materials}
- G01N 2030/8836: {involving saccharides}
- Y10T 436/143333: Saccharide [e.g., DNA, etc.]
- C08B: POLYSACCHARIDES; DERIVATIVES THEREOF (polysaccharides containing less than six saccharide radicals attached to each other by glycosidic linkages C07H; fermentation or enzyme-using processes C12P19/00; sugar industry C13; production of cellulose D21)
- 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 to the detection and quantitation of cyclodextrins and cyclodextrin derivatives in solutions comprising a protein. The invention further relates to methods of evaluating pharmaceutical preparations for the presence of residual cyclodextrins.

## Claims

### Claim 1

A method for recovering a cyclodextrin or cyclodextrin derivative from a solution comprising a protein recombinantly expressed in the presence of the cyclodextrin or cyclodextrin derivative, the method comprising loading the solution on a size exclusion chromatography (SEC) column, wherein the solution does not undergo a step of extraction prior to being loaded on the SEC column; separating the cyclodextrin or cyclodextrin derivative from the recombinantly expressed protein by SEC; contacting the cyclodextrin or cyclodextrin derivative with an agent that forms a detectable inclusion complex with the separated cyclodextrin or cyclodextrin derivative; and measuring the amount of a signal from the inclusion complex, wherein the signal is indicative of the quantity of the cyclodextrin or cyclodextrin derivative recovered from the solution.

### Claim 2 (depends on 1)

The method of claim 1, wherein the cyclodextrin or cyclodextrin derivative is methyl-β-cyclodextrin.

### Claim 3 (depends on 1)

The method of claim 1, wherein the recombinantly expressed protein is an antibody.

### Claim 4 (depends on 1)

The method of claim 1, wherein the solution is a pharmaceutical preparation.

### Claim 5 (depends on 1)

The method of claim 1, wherein the solution comprises less than 20 μg/mL of the cyclodextrin or cyclodextrin derivative.

### Claim 6 (depends on 1)

The method of claim 1, wherein the agent is a fluorescent compound.

### Claim 7 (depends on 6)

The method of claim 6, wherein the agent is 1-naphthol.

### Claim 8 (depends on 1)

The method of claim 1, wherein the method has a cyclodextrin or a cyclodextrin derivative limit of detection of less than 10 μg/mL.

### Claim 9 (depends on 8)

The method of claim 8, wherein the method has a cyclodextrin or a cyclodextrin derivative limit of detection of less than 2 μg/mL.

### Claim 10 (depends on 1)

The method of claim 1, wherein the SEC column has a separation range of about 5-20 kDa to about 75-300 kDa.

### Claim 11 (depends on 1)

The method of claim 1, wherein the SEC column has a particle size of about 2 to about 6 microns.

### Claim 12 (depends on 11)

The method of claim 11, wherein the SEC column has a particle size of about 4 microns.

### Claim 13 (depends on 1)

The method of claim 1, wherein at least 97%±5% of the cyclodextrin or cyclodextrin derivative is recovered.

### Claim 14 (depends on 1)

The method of claim 1, wherein the separation uses a mobile phase comprising a salt at a concentration of about 50 mM to about 300 mM.

### Claim 15 (depends on 14)

The method of claim 14, wherein the solution and the mobile phase have a pH of about 2.0 to about 8.0.

### Claim 16 (depends on 15)

The method of claim 15, wherein the solution and the mobile phase have a pH of about 5.0 to about 7.5.

### Claim 17 (depends on 14)

The method of claim 14, wherein the agent is present in the mobile phase during the separating step.

### Claim 18 (depends on 1)

The method of claim 1, wherein the recombinantly expressed protein is at least 90% pure.

### Claim 19 (depends on 18)

The method of claim 18, wherein the recombinantly expressed protein is at least 95% pure.

### Claim 20 (depends on 19)

The method of claim 19, wherein the recombinantly expressed protein is at least 98% pure.