# US 2013/0153420 A1 · Microfluidic Detection of Analytes

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

- Number: US 2013/0153420 A1
- Document ID: US20130153420A1
- Status: Published application
- Kind: A1
- Application number: 13/536,423
- Filed: 2012-06-28
- Published: 2013-06-20
- Earliest claimed: 2005-10-04
- Assignees: Headway Technologies, Inc.
- Inventors: Celine Hu
- Web page: https://patentgrep.com/patent/US20130153420A1

## CPC

- G01N 27/447: using electrophoresis
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- G01N 1/40: Concentrating samples
- G01N 33/536: with immune complex formed in liquid phase
- B01L 2200/027: for microfluidic devices
- B01L 2300/0636: Integrated biosensor, microarrays
- B01L 2400/0421: electrophoretic flow
- B33Y 80/00: Products made by additive manufacturing
- G01N 2001/4038: {electric methods, e.g. electromigration, electrophoresis, ionisation}
- 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
- B33Y: ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING

## Abstract

An apparatus and methods for concentrating samples for application to microfluidic devices are disclosed. The methods involve electrophoresing charged molecules from a high volume sample into a smaller volume. The analyte of interest can be a charged molecule or can be modified to be charged using, for example, one or more ionic moieties.

## Claims

### Claim at position 1

1-55. (canceled)

### Claim 56

A method for introducing an analyte of interest in a sample to a microfluidic device, comprising:
providing an aqueous sample in a large volume reservoir, said sample containing the analyte and having a volume of greater than or equal to 1 microliter, wherein the analyte is charged or has been associated with a charged molecule;
electrophoresing the analyte from the large volume reservoir to a microfluidic device via a connector, wherein the microfluidic device comprises an analyis area, and the large volume reservoir and the microfluidic device are fluidically connected via the connector; and
continuously flowing the analyte to the analysis area for a time sufficient to result in a higher concentration of the analyte in the analysis area than in the sample.

### Claim 57 (depends on 56)

The method of claim 56, wherein said sample has a volume of from 1 microliter to 100 milliliters.

### Claim 58 (depends on 57)

The method of claim 57, wherein said sample has a volume of from 1 microliter to 20 mililiters.

### Claim 59 (depends on 56)

The method of claim 56, further comprising providing at least one other aqueous sample in a large volume reservoir fluidically connected to the microfluidic device, the microfluidic device further comprising at least one other analysis area for said one other sample.

### Claim 60 (depends on 56)

The method of claim 56, wherein the large volume reservoir is a chamber that is not integral with said microfluidic device.

### Claim 61 (depends on 60)

The method of claim 60, wherein said large volume reservoir is a well of a microwell plate, an eppendorf tube, or a test tube.

### Claim 62 (depends on 56)

The method of claim 56, wherein said connector is a capillary tube,

### Claim 63 (depends on 56)

The method of claim 56, wherein said analyte is covalently bound to an ionic moiety or is associated with a charged molecule by being bound to a carrier molecule that is ionically charged or is modified to be ionically charged.

### Claim 64 (depends on 63)

The method of claim 63, wherein the carrier molecule is selected from the group consisting of an antibody, a receptor, a ligand, and an antigen.

### Claim 65 (depends on 56)

The method of claim 56, wherein the analysis area comprises a capture site comprising a capture agent, said method further comprising transporting the analyte across the capture site under conditions where the analyte is captured by the capture agent.

### Claim 66 (depends on 65)

The method of claim 65, wherein said transporting across the capture site is by electrophoresis.

### Claim 67 (depends on 65)

The method of claim 65, further comprising detecting the analyte bound by the capture agent.

### Claim 68 (depends on 56)

The method of claim 56 further comprising immobilizing, said analyte of interest on a microparticle before electrophoresis.

### Claim 69 (depends on 68)

The method of claim 68, wherein said microparticle is a magnetic microparticle.

### Claim 70 (depends on 68)

The method of claim 68, wherein said microparticle is coated with at least one receptor, antibody, or anti-ligand that specifically binds said analyte of interest or a group of molecules including said analyte of interest.

### Claim 71 (depends on 70)

The method of claim 70, wherein the method further comprises detecting a label on a second antibody hound to said analyte of interest or said group of molecules including said analyte of interest.

### Claim 72 (depends on 68)

The method of claim 68, further comprising the step of removing the analyte from the microparticle prior to electrophoresis.

### Claim 73

A method for introducing an analyte of interest in a sample to a microfluidic device, comprising:
providing an aqueous sample in a large volume reservoir comprising a first microelectrode operably attached thereto, said sample containing the analyte and having a volume of greater than or equal to 1 microliter, wherein the analyte of interest is charged or has been associated with a charged molecule;
electrophoresing the analyte via a connector from the large volume reservoir to a staging reservoir positioned in a microfluidic device, wherein the connector fluidically connects the large volume reservoir and the staging reservoir, and wherein the microfluidic device comprises an analysis area, a second microelectrode located in the staging reservoir and a third microelectrode located in or distal to at least a portion of the analysis area, said microelectrodes being configured such that an electric field may be generated between the electrodes and a charged analyte electrophoresed from the first microelectrode to the second microelectrode and from the second microelectrode to the third microelectrode; and
transporting the analyte to the analysis area for a time sufficient to result in a higher concentration of analyte in the analysis area than in the sample.

### Claim 74 (depends on 73)

The method of claim 73, wherein the aqueous sample in the large volume reservoir has a volume which is about 100 to about 1000 times greater than the volume of the staging reservoir.

### Claim 75 (depends on 73)

The method of claim 73, wherein said sample has a volume of from 1 microliter to 100 mililiters.

### Claim 76 (depends on 75)

The method of claim 75, wherein said sample has a volume of from 1 microliter to 20 mililiters.

### Claim 77

A method for introducing an analyte of interest to a microfluidic device, comprising:
introducing a solid phase-bound analyte into a large volume reservoir to provide a sample, said sample having a volume of greater than or equal to 1 microliter, wherein the analyte is charged or has been associated with a charged molecule;
exposing the solid support-bound analyte to conditions sufficient to free the analyte from the solid support;
electrophoresing the analyte from the large volume reservoir to a microfluidic device via a connector, wherein the microfluidic device comprises an analysis area, and the large volume reservoir and the microfluidic device are fluidically connected via the connector; and
transporting the analyte to the analysis area for a time sufficient to result in a higher concentration of analyte in the analysis area than in the sample, wherein the analyte is continuously flowed from the large volume reservoir to the analysis area during said time.

### Claim 78 (depends on 77)

The method of claim 77, wherein said sample has a volume of from 1 microliter to 100 mililiters.

### Claim 79 (depends on 78)

The method of claim 78, wherein said sample has a volume of from 1 microliter to 20 mililiters.

### Claim 80 (depends on 77)

The method of claim 77 wherein the solid phase is a microparticle,

### Claim 81 (depends on 80)

The method of claim 80, wherein said microparticle is a magnetic microparticle.