# US 2014/0154703 A1 · Circulating tumor cell capture on a microfluidic chip incorporating both affinity and size

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- Number: US 2014/0154703 A1
- Document ID: US20140154703A1
- Status: Published application
- Kind: A1
- Application number: 13/978,123
- Filed: 2012-01-06
- Published: 2014-06-05
- Earliest claimed: 2011-01-06
- Inventors: Alison Skelley; Denis Smirnov; Yi Dong; Keith D. Merdek; Kam Sprott; Walter Carney; Chunsheng Jiang; Richard Huang; Ioana Lupascu
- Web page: https://patentgrep.com/patent/US20140154703A1

## CPC

- G01N 33/575: for cancer
- B01L 3/502761: {specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules}
- B01L 2200/0652: Sorting or classification of particles or molecules
- B01L 2200/0668: Trapping microscopic beads
- B01L 2300/0822: Slides
- B01L 2300/0877: Flow chambers
- B01L 2300/168: Specific optical properties, e.g. reflective coatings
- B01L 2400/086: using baffles or other fixed flow obstructions
- G01N 30/6095: {Micromachined or nanomachined, e.g. micro- or nanosize}
- 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)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

## Abstract

The invention encompasses methods and devices for diagnosing, theranosing, or prognosing a condition in a patient by enriching a sample in rare cells or other particles. The devices can be a microfluidic device comprising an array of obstacles and one or more binding moieties. The devices and methods can allow for enrichment of cells based on size and affinity, recovery of cells or other particles in locations on the microfluidic device, release of cells or other particles from the microfluidic device, flow of sample through the microfluidic device, and retention of rare cells or other particles from a sample obtained from a patient having a condition.

## Claims

### Claim at position 1

-53. (canceled)

### Claim 54

A microfluidic device comprising an array of obstacles coated with antibodies wherein a surface of the device is functionalized with dextran or dextran derivatives.

### Claim 55

A microfluidic device comprising an array of obstacles coated with antibodies wherein a surface of the device has a contact angle of less than 15° over at least 10 hours.

### Claim at position 4

-71. (canceled)

### Claim 72

A method for capture and release of cells or cell fragments of interest, the method comprising: (a) flowing a sample comprising cells or cell fragments of interest on a surface coated with carbohydrate and binding moiety that selectively binds a cell surface marker selectively present on the cells or cell fragments of interest, and (b) using an enzyme that selectively cleaves the carbohydrate or a biotin derivative wherein the biotin derivative competitively releases biotin conjugates, or both, to thereby release the cells or cell fragments of interest from the surface.

### Claim 73 (depends on 72)

The method according to claim 72, wherein a biotin derivative that competitively releases biotin conjugates comprises desthiobiotin.

### Claim 74 (depends on 72)

The method according to claim 72, wherein an enzyme that selectively cleaves the carbohydrate comprises dextranase, a glycosyltransferase, a glycoside hydrolase, a transglycosidase, a phosphorylase, or a lyase.

### Claim at position 8

-78. (canceled)

### Claim 79 (depends on 54)

The device of claim 54, wherein the device is capable of capturing at least 60% of circulating tumor cells in a sample when a concentration of the antibodies in a solution that coat the array of obstacles is between about 10 μg/mL to about 100 μg/mL.

### Claim 80 (depends on 54)

The device of claim 54, wherein a concentration of the dextran or the dextran derivatives in a solution used to functionalize the surface of the device is from 0.01% to 5% (w/w) or from 0.05% to 2% (w/w).

### Claim 81 (depends on 54)

The device of claim 54, wherein the antibodies coated on the array of obstacles are capable of selectively capturing cells, cell fragments, particles, or any combination thereof.

### Claim 82 (depends on 81)

The device of claim 81, wherein the cells comprise epithelial cells, non-epithelial cells, non-epithelial tumor cells, cells undergoing epithelial to mesenchymal transition, cancer stem cells, mesenchymal cells, or any combination thereof.

### Claim 83 (depends on 81)

The device of claim 81, wherein the cell fragments comprise proteins or nucleic acids.

### Claim 84 (depends on 55)

The device of claim 55, wherein the surface of the device is functionalized with a carbohydrate.

### Claim 85 (depends on 84)

The device of claim 84, wherein the carbohydrate comprises dextran, chitin, chitosan, alginate, cellulose, methylcellulose, starch, heparin, agarose, concanavalin A, callose, laminarin, chrysolaminarin, xylan, arabinoxylan, mannan, fucoidan, galactomannan, or any combination thereof.

### Claim 86 (depends on 84)

The device of claim 84, wherein a concentration of the carbohydrate in a solution used to functionalize the surface of the device is from 0.01% to 5% (w/w), 0.01% to 4% (w/w), 0.01% to 3.75% (w/w), 0.01% to 3.5% (w/w), 0.01% to 3.25% (w/w), 0.01% to 3% (w/w), 0.01% to 2.75% (w/w), 0.01% to 2.5% (w/w), 0.01% to 2.25% (w/w), 0.01% to 2% (w/w), 0.01% to 1.75% (w/w), 0.01% to 1.5% (w/w), 0.01% to 1.25% (w/w), 0.01% to 1% (w/w), 0.01% to 0.75% (w/w), 0.01% to 0.5% (w/w), 0.01% to 0.25% (w/w), 0.05% to 2% (w/w), 0.05% to 1.9% (w/w), 0.05% to 1.8% (w/w), 0.05% to 1.7% (w/w), 0.05% to 1.6% (w/w), 0.05% to 1.5% (w/w), 0.05% to 1.4% (w/w), 0.05% to 1.3% (w/w), 0.05% to 1.2% (w/w), 0.05% to 1.1% (w/w), 0.05% to 1% (w/w), 0.05% to 0.9% (w/w), 0.05% to 0.8% (w/w), 0.05% to 0.7% (w/w), 0.05% to 0.6% (w/w), 0.05% to 0.5% (w/w), 0.05% to 0.4% (w/w), 0.05% to 0.3% (w/w), 0.05% to 0.2% (w/w), or 0.05% to 0.1% (w/w).

### Claim 87 (depends on 72)

The method according to claim 72, wherein the binding moiety is covalently bonded to the carbohydrate.

### Claim 88 (depends on 72)

The method according to claim 72, wherein the binding moiety is bonded to the carbohydrate via a linker.

### Claim 89 (depends on 72)

The method according to claim 72, wherein the binding moiety is an affinity tagged ligand.

### Claim 90 (depends on 72)

The method according to claim 72, wherein the binding moiety is selected from the group consisting of: avidin, NeutrAvidin, StreptAvidin, or CaptAvidin.

### Claim 91 (depends on 72)

The method according to claim 72, wherein the binding moiety enables capture of epithelial cells, non-epithelial cells, non-epithelial tumor cells, cells undergoing epithelial to mesenchymal transition, cancer stem cells, mesenchymal cells, cellular fragments, proteins, nucleic acids particles, or any combination thereof.

### Claim 92 (depends on 72)

The method according to claim 72, wherein the carbohydrate is dextran, chitin, chitosan, alginate, cellulose, methylcellulose, starch, heparin, agarose, concanavalin A, callose, laminarin, chrysolaminarin, xylan, arabinoxylan, mannan, fucoidan, galactomannan, or any combination thereof.

### Claim 93 (depends on 92)

The method according to claim 92, wherein a concentration of the carbohydrate in solution that is dextran used to coat the surface is from 0.01% to 5% (w/w) or from 0.05% to 2% (w/w).