# US 12,216,117 B2 · Reagent storage in an assay device

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- Number: US 12,216,117 B2
- Document ID: US12216117B2
- Status: Granted
- Kind: B2
- Application number: 17/553,441
- Filed: 2021-12-16
- Published: 2022-08-11
- Granted: 2025-02-04
- Earliest claimed: 2010-10-14
- Assignees: Meso Scale Technologies, LLC.
- Applicants: Meso Scale Technologies, LLC.
- Inventors: Sudeep Kumar; George Sigal; Michael Tsionsky
- Web page: https://patentgrep.com/patent/US12216117B2

## CPC

- G01N 33/54386: {Analytical elements}
- C12Q 1/6823: Release of bound markers
- C12Q 1/6832: Enhancement of hybridisation reaction
- C12Q 1/6834: Enzymatic or biochemical coupling of nucleic acids to a solid phase
- G01N 33/5306: {Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding}
- G01N 2446/00: Magnetic particle immunoreagent carriers
- G01N 2458/30: Electrochemically active labels
- 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)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The invention relates to methods for conducting binding assays in an assay device that includes one or more storage and use zone. The storage zones of the assay device are configured to house one or more reagents used in an assay conducted in the use zone of the device.

## Claims

### Claim 1

An assay device comprising one or more reagent spaces and one or more use zones:
(a) said one or more reagent spaces comprise a storage zone comprising more than one storage surface to which two or more surface-reagent complexes are bound and thereby confined to said storage zone, said two or more surface-reagent complexes comprising a first surface-reagent complex and a second surface-reagent complex, said first surface-reagent complex comprising
(i) a first reagent linked to a first targeting agent;
(ii) a first surface linked to a second targeting agent:
and said second surface-reagent complex comprising:
(iii) a second reagent linked to a third targeting agent; and
(iv) a second surface linked to a fourth targeting agent,
wherein said first reagent and said first surface and said second reagent and said second surface are linked, respectively, in said first and second surface-reagent complexes, via a selective releasable binding interaction between said first and second targeting agents and said third and fourth targeting agents, respectively, wherein said first reagent is released from said first surface-reagent complex by a first set of conditions that differs from a second set of conditions used to release said second reagent from said second surface-reagent complex; and
(b) said one or more use zones are configured to use at least one of said first and second reagents in an assay for an analyte of interest in a sample.

### Claim 2 (depends on 1)

The assay device of claim 1 wherein said one or more use zones each comprise an additional reagent used in said assay.

### Claim 3 (depends on 2)

The assay device of claim 2 wherein said additional reagent is a binding reagent that binds a component of said assay conducted in said use zone.

### Claim 4 (depends on 3)

The assay device of claim 3 wherein said binding reagent binds said analyte.

### Claim 5 (depends on 3)

The assay device of claim 3 wherein said binding reagent binds a complex formed between said additional reagent and said analyte.

### Claim 6 (depends on 5)

The assay device of claim 5 wherein said one or more use zones each comprise a solid support and said additional reagent is bound to said solid support.

### Claim 7 (depends on 2)

The assay device of claim 2 wherein said additional reagent binds said analyte.

### Claim 8 (depends on 2)

The assay device of claim 2 wherein said additional reagent comprises a detectable label.

### Claim 9 (depends on 8)

The assay device of claim 8 wherein said detectable label is an ECL label.

### Claim 10 (depends on 2)

The assay device of claim 2 wherein said storage zone and said one or more use zones are in fluidic communication along a fluid path.

### Claim 11 (depends on 10)

The assay device of claim 10 wherein said one or more use zones each comprise two or more assay regions each configured to use said additional reagent in one or more assays conducted with said sample in said assay device.

### Claim 12 (depends on 11)

The assay device of claim 11 wherein a first assay region of said one or more use zones is configured to conduct an assay for a first analyte of interest in said sample and an additional assay region in said one or more use zones is configured to conduct an assay for an additional analyte of interest in said sample.

### Claim 13 (depends on 11)

The assay device of claim 11 wherein a first assay region of said one or more use zones is configured to conduct a first assay for said analyte of interest in said sample and an additional assay region of said one or more use zones is configured to conduct a second assay for said analyte of interest in said sample.

### Claim 14 (depends on 11)

The assay device of claim 11 wherein each of said two or more assay regions comprise an additional reagent used in said assay.

### Claim 15 (depends on 14)

The assay device of claim 14 wherein said additional reagent is an additional binding reagent.

### Claim 16 (depends on 1)

The assay device of claim 1 wherein said first and second surfaces are particles.

### Claim 17 (depends on 1)

The assay device of claim 1 wherein said first and second surfaces are roughened such that the surface area accessible to a component capable of binding to said first and second surface is at least three-fold larger than the surface area of a flat surface.

### Claim 18 (depends on 1)

The assay device of claim 1 wherein said first and second surfaces are roughened such that the surface area accessible to a component capable of binding to said first and second surfaces is at least two-fold larger than the surface area of a flat surface.

### Claim 19 (depends on 1)

The assay device of claim 1 wherein said first and second surfaces comprise a composite material including exposed particles distributed in a matrix.

### Claim 20 (depends on 19)

The assay device of claim 19 wherein said composite material comprises carbon particles, graphitic particles, or carbon nanotubes.

### Claim 21 (depends on 19)

The assay device of claim 19 wherein said composite is etched.

### Claim 22 (depends on 19)

The assay device of claim 19 wherein said first and second surfaces comprise one or more indentations or raised features.

### Claim 23 (depends on 1)

The assay device of claim 1 wherein said first and second surfaces comprise a hydrogel.

### Claim 24 (depends on 1)

The assay device of claim 1 wherein each reagent is released from each surface-reagent complex by subjecting said storage zone to increased or decreased temperature, pH changes, an electric potential, a change in ionic strength, competition, and combinations thereof.

### Claim 25 (depends on 24)

The assay device of claim 24 wherein each reagent is released by subjecting said storage zone to increased temperature.

### Claim 26 (depends on 25)

The assay device of claim 25 wherein said increased temperature exceeds the melting temperature of said binding interaction.

### Claim 27 (depends on 1)

The assay device of claim 1 wherein said assay device is a cartridge.

### Claim 28 (depends on 1)

The assay device of claim 1 wherein said assay device is a multi-well assay plate and said use zone is positioned within a well of said assay plate.

### Claim 29 (depends on 1)

The assay device of claim 1 wherein said storage zone is located on a supplemental surface of said well that does not overlap with said use zone.

### Claim 30 (depends on 1)

The assay device of claim 1 wherein said device is configured to conduct a multiplexed measurement.

### Claim 31

A multi-well assay plate comprising a plate body comprising a plurality of assay wells and one or more reagent spaces defined in said plate body, wherein the one or more reagent spaces are in fluidic communication with one or more surrounding wells of said plurality of wells, wherein
(a) said one or more reagent spaces comprise a storage zone comprising a storage surface to which two or more surface-reagent complexes are bound and thereby confined to said storage zone, said two or more surface-reagent complexes comprising a first surface-reagent complex and a second surface-reagent complex, said first surface-reagent complex comprising:
(i) a first reagent linked to a first targeting agent; and
(ii) a first surface linked to a second targeting agent, and said second surface-reagent complex comprising:
(iii) a second reagent linked to a third targeting agent; and
(iv) a second surface linked to a fourth targeting agent,
wherein said first reagent and said first surface and said second reagent and said second surface are linked, respectively, in said first and second surface-reagent complexes, via a selective releasable binding interaction between said first and second targeting agents and said third and fourth targeting agents, respectively, wherein said first reagent is released from said first surface-reagent complex by a first set of conditions that differ from a second set of conditions used to release said second reagent from said second surface-reagent complex; and
(b) said one or more surrounding wells are configured to use at least one of said first and second reagents in an assay for an analyte of interest in a sample.

### Claim 32 (depends on 31)

The multi-well assay plate of claim 31 wherein said one or more surrounding wells each comprise an additional reagent used in said assay.

### Claim 33 (depends on 31)

The multi-well assay plate of claim 31 wherein at least one of said first and second reagents comprises a detectable label.

### Claim 34 (depends on 31)

The multi-well assay plate of claim 31 wherein said one or more surrounding wells each comprise two or more assay regions each configured to use said at least one of said first and second reagent in one or more assays conducted with said sample in said assay device.

### Claim 35 (depends on 31)

The multi-well assay plate of claim 31 wherein said first and second surfaces are particles.

### Claim 36 (depends on 31)

The multi-well assay plate of claim 31 wherein said first and second surfaces are roughened such that the surface area accessible to a component capable of binding to said first and second surface is at least three-fold larger than the surface area of a flat surface.

### Claim 37 (depends on 31)

The multi-well assay plate of claim 31 wherein said first and second surfaces are roughened such that the surface area accessible to a component capable of binding to said first and second surface is at least two-fold larger than the surface area of a flat surface.

### Claim 38 (depends on 31)

The multi-well assay plate of claim 31 wherein said first and second surfaces comprise a composite material including exposed particles distributed in a matrix.

### Claim 39 (depends on 31)

The multi-well assay plate of claim 31 wherein said first and second surfaces comprise a hydrogel.

### Claim 40 (depends on 31)

The multi-well assay plate of claim 31 wherein said first reagent is released by subjecting said storage zone to a first set of conditions selected from the group consisting of increased or decreased temperature, pH change, application of an electrical potential, change in ionic strength, competition and combinations thereof and said second reagent is released by subjecting said storage zone to a second set of conditions selected from the group consisting of increased or decreased temperature, pH change, application of an electrical potential, change in ionic strength, competition and combinations thereof, wherein said first and second set of conditions are different.

### Claim 41 (depends on 31)

The multi-well assay plate of claim 31 wherein said device is configured to conduct a multiplexed measurement.

### Claim 42

A multi-well assay plate comprising a plate body comprising (a) a plurality of assay wells comprising one or more use zones; and (b) one or more storage surfaces positioned in said plate body, wherein the one or more storage surfaces do not overlap with said one or more use zones, wherein the one or more storage surfaces are in fluidic communication with one or more wells of said plurality of wells, wherein
(a) said one or more storage surfaces comprise two or more surface-reagent complexes bound to said one or more storage surfaces,
said two or more surface-reagent complexes comprising a first surface-reagent complex and a second surface-reagent complex, said first surface-reagent complex comprising:
(i) a first reagent linked to a first targeting agent; and
(ii) a first surface linked to a second targeting agent, and said second surface-reagent complex comprising:
(iii) a second reagent linked to a third targeting agent; and
(iv) a second surface linked to a fourth targeting agent,
wherein said first reagent and said first surface and said second reagent and said second surface are linked, respectively, in said first and second surface-reagent complexes, via a selective releasable binding interaction between said first and second targeting agents and said third and fourth targeting agents, respectively, wherein said first reagent is released from said first surface-reagent complex by a first set of conditions that differ from a second set of conditions used to release said second reagent from said second surface-reagent complex; and
(b) said one or more use zones are each configured to use at least one of said first and second reagents in an assay for one or more analytes of interest in a sample.

### Claim 43 (depends on 42)

The multi-well assay plate of claim 42 wherein said one or more use zones each comprise an additional reagent used in said assay.

### Claim 44 (depends on 42)

The multi-well assay plate of claim 42 wherein at least one of said first and second reagents is a binding reagent that binds a component of said assay conducted in said use zone.

### Claim 45 (depends on 42)

The multi-well assay plate of claim 42 wherein at least one of said first and second reagents comprise a detectable label.

### Claim 46 (depends on 42)

The multi-well assay plate of claim 42 wherein said one or more use zones each comprise two or more assay regions each configured to use at least one of said first and second reagent in one or more assays conducted with said sample in said assay device.

### Claim 47 (depends on 42)

The multi-well assay plate of claim 42 wherein said first and second surfaces are particles.

### Claim 48 (depends on 42)

The multi-well assay plate of claim 42 wherein said first and second surfaces are roughened such that the surface area accessible to a component capable of binding to said first and second surface is at least three-fold larger than the surface area of a flat surface.

### Claim 49 (depends on 42)

The multi-well assay plate of claim 42 wherein said first and second surfaces are roughened such that the surface area accessible to a component capable of binding to said first and second surface is at least two-fold larger than the surface area of a flat surface.

### Claim 50 (depends on 42)

The multi-well assay plate of claim 42 wherein said first and second surfaces comprise a composite material including exposed particles distributed in a matrix.

### Claim 51 (depends on 42)

The multi-well assay plate of claim 42 wherein said first and second surfaces comprise a hydrogel.

### Claim 52 (depends on 42)

The multi-well assay plate of claim 42 wherein said first reagent is released by subjecting said storage zone to a first set of conditions selected from the group consisting of increased or decreased temperature, pH change, application of an electrical potential, change in ionic strength, competition and combinations thereof and said second reagent is released by subjecting said storage zone to a second set of conditions selected from the group consisting of increased or decreased temperature, pH change, application of an electrical potential, change in ionic strength, competition and combinations thereof, wherein said first and second set of conditions are different.

### Claim 53 (depends on 42)

The multi-well assay plate of claim 42 wherein said storage zone is located on a supplemental surface of said well that does not overlap with said use zone.

### Claim 54 (depends on 42)

The multi-well assay plate of claim 42 wherein said device is configured to conduct a multiplexed measurement.