US5637469A — Methods and apparatus for the detection of an ana…

Claim 1

Independent Claim 134 words

A device for detecting the presence of an analyte in a fluid sample, the device comprising: a solid substrate microfabricated to define: a sample inlet port, said port being dimensioned for connection to a pump means; and a mesoscale flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds to analyte in said fluid sample, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting binding of said analyte to said binding moiety.

Claim 2

Independent Claim 134 words

A device for detecting the presence of an analyte in a fluid sample, the device comprising: a solid substrate microfabricated to define: a sample inlet port, said port being dimensioned for connection to a pump means; and a mesoscale flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds to analyte in said fluid sample, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting binding of said analyte to said binding moiety.

Claim 3

Dependent Claim 22 words

The device of claim 1, wherein said binding moiety is immobilized on a surface of a solid particle in said binding region.

Claim 4

Dependent Claim 17 words

The device of claim 1, wherein said analyte and said immobilized binding moiety comprise a ligand/receptor pair.

Claim 5

Dependent Claim 70 words

The device of claim 1, wherein said analyte is an intracellular molecular component in a cell-containing liquid biological sample, said device further comprising: cell lysing means in said mesoscale flow system in fluid communication with said flow channel, said analyte binding region being downstream from said cell lysing means; and means for engaging cells in said cell-containing sample with said cell lysing means, thereby to release said intracellular molecular component.

Claim 6

Dependent Claim 26 words

The device of claim 4, wherein said cell lysing means comprises a portion of said flow channel having cell-membrane piercing protrusions extending from a wall thereof.

Claim 7

Dependent Claim 29 words

The device of claim 4, wherein said cell lysing means comprises a region of restricted cross-sectional dimension sufficient to permit passage of intracellular molecules while prohibiting passage of cells.

Claim 8

Dependent Claim 57 words

The device of claim 4, wherein said analyte is a cell population in said sample, said device further comprising: a cell separation region, upstream from said analyte binding region comprising an immobilized cell binding moiety that binds cell surface molecules of said cell population; and pump means for inducing flow of said sample to said separation region.

Claim 9

Dependent Claim 12 words

The device of claim 1, wherein said solid substrate comprises microfabricated silicon.

Claim 10

Dependent Claim 32 words

The device of claim 1, wherein said sample flow channel and said binding region are microfabricated in a surface of said solid substrate and enclosed by a cover adhered to said surface.

Claim 11

Dependent Claim 31 words

The device of claim 1, wherein said binding means comprises a window for optically probing said analyte binding region, said window being disposed over said analyte binding region on said substrate.

Claim 12

Dependent Claim 38 words

The device of claim 10, wherein said analyte binding region comprises particles having said analyte specific binding moiety immobilized on the surface thereof which, in the presence of an analyte, induce particle agglomeration optically detectable through said window.

Claim 13

Dependent Claim 54 words

The device of claim 10, wherein said substrate further defines a control region in fluid communication with said sample inlet port and a control region window, disposed over said control region on said substrate, for optically probing said control region whereby data determined optically in said control and analyte binding regions may be compared.

Claim 14

Dependent Claim 47 words

The device of claim 10, wherein said flow system further comprises: a fractal region in fluid communication with said flow channel comprising bifurcations leading to plural secondary flow channels; and pump means for inducing flow of a biological sample through said flow channel and said fractal region.

Claim 15

Dependent Claim 22 words

The device of claim 1, wherein the width and depth of said flow channel each are between 2.0 um and 500 um.

Claim 16

Independent Claim 161 words

A device for detecting the presence of an analyte in a fluid sample, the device comprising: a solid substrate microfabricated to define: a sample inlet port; and a mesoscale flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety immobilized on a surface of said analyte binding region, wherein said binding moiety specifically binds to analyte in said sample, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; a pump means for delivering to said analyte binding region a labelled substance which binds analyte bound to said binding moiety to produce an optically detectable signal indicative of the presence of said analyte; and a binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting said optically detectable signal, thereby to determine the presence of the analyte.

Claim 17

Independent Claim 161 words

A device for detecting the presence of an analyte in a fluid sample, the device comprising: a solid substrate microfabricated to define: a sample inlet port; and a mesoscale flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety immobilized on a surface of said analyte binding region, wherein said binding moiety specifically binds to analyte in said sample, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; a pump means for delivering to said analyte binding region a labelled substance which binds analyte bound to said binding moiety to produce an optically detectable signal indicative of the presence of said analyte; and a binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting said optically detectable signal, thereby to determine the presence of the analyte.

Claim 18

Dependent Claim 50 words

The device of claim 15, wherein said binding means comprises: a window for optically probing said analyte binding region, said window being disposed over said analyte binding region on said substrate; and optical binding means disposed over said window for detecting the presence of said detectable signal through said window.

Claim 19

Dependent Claim 14 words

The device of claim 15, wherein said optically detectable signal is a luminescent signal.

Claim 20

Dependent Claim 14 words

The device of claim 15, wherein said optically detectable signal is a fluorescent signal.

Claim 21

Independent Claim 162 words

A device for detecting the presence of multiple analytes in a fluid sample, the device comprising: a solid substrate microfabricated to define: a sample inlet port; and a first and at least a second mesoscale flow system, each said flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds to analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a binding means associated with each analyte binding region or in fluid communication with each analyte binding region for detecting the binding of analyte to the binding moiety in each analyte binding region, thereby to determine the presence of said analyte; and wherein the analyte binding regions of said flow systems comprise different immobilized binding moieties capable of binding different analytes.

Claim 22

Independent Claim 162 words

A device for detecting the presence of multiple analytes in a fluid sample, the device comprising: a solid substrate microfabricated to define: a sample inlet port; and a first and at least a second mesoscale flow system, each said flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds to analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a binding means associated with each analyte binding region or in fluid communication with each analyte binding region for detecting the binding of analyte to the binding moiety in each analyte binding region, thereby to determine the presence of said analyte; and wherein the analyte binding regions of said flow systems comprise different immobilized binding moieties capable of binding different analytes.

Claim 23

Independent Claim 149 words

A device for detecting the presence of an analyte in a fluid sample, the device comprising: a solid substrate microfabricated to define: a sample inlet port; and a mesoscale flow system comprising: a tortuous sample flow channel extending from said inlet port, wherein said tortuous flow channel is microfabricated with a length which allows timed mixing of reagents and sample fluid; and an analyte binding region in fluid communication with said tortuous flow channel comprising a binding moiety immobilized on a surface in said analyte binding region, said binding moiety specifically binds to said analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting the binding of analyte to the binding moiety thereby to determine the presence of the analyte.

Claim 24

Independent Claim 149 words

A device for detecting the presence of an analyte in a fluid sample, the device comprising: a solid substrate microfabricated to define: a sample inlet port; and a mesoscale flow system comprising: a tortuous sample flow channel extending from said inlet port, wherein said tortuous flow channel is microfabricated with a length which allows timed mixing of reagents and sample fluid; and an analyte binding region in fluid communication with said tortuous flow channel comprising a binding moiety immobilized on a surface in said analyte binding region, said binding moiety specifically binds to said analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting the binding of analyte to the binding moiety thereby to determine the presence of the analyte.

Claim 25

Independent Claim 202 words

A method for detecting the presence of an analyte in a fluid sample, the method comprising the steps of: (i) providing a device comprising: a solid substrate microfabricated to define: a sample inlet port, said port being dimensioned for connection to a pump means; and a mesoscale flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds to said analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting binding of said analyte to said binding moiety, thereby to determine the presence of said analyte; (ii) delivering a sample to said inlet port and, by a pump means, through said flow system to said analyte binding region to permit binding of said analyte to said binding moiety; and (iii) detecting the binding of analyte to said binding moiety with said binding means, thereby to determine the presence of the analyte.

Claim 26

Independent Claim 202 words

A method for detecting the presence of an analyte in a fluid sample, the method comprising the steps of: (i) providing a device comprising: a solid substrate microfabricated to define: a sample inlet port, said port being dimensioned for connection to a pump means; and a mesoscale flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds to said analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting binding of said analyte to said binding moiety, thereby to determine the presence of said analyte; (ii) delivering a sample to said inlet port and, by a pump means, through said flow system to said analyte binding region to permit binding of said analyte to said binding moiety; and (iii) detecting the binding of analyte to said binding moiety with said binding means, thereby to determine the presence of the analyte.

Claim 27

Dependent Claim 44 words

The method of claim 21, wherein said binding means in the device provided in step (i) comprises a window disposed over said analyte binding region on said substrate; and wherein step (iii) includes the step of optically probing said binding region through said window.

Claim 28

Dependent Claim 77 words

The method of claim 22, wherein said substrate provided in step (i) further defines a control region in fluid communication with said sample inlet port and a control region window, disposed over said control region on said substrate, for optically probing said control region whereby data determined optically in said control and analyte binding regions may be compared; and wherein step (iii) includes the step of optically probing and comparing said control region and said binding region.

Claim 29

Dependent Claim 106 words

The method of claim 21, wherein the device provided in step (i) further includes: a pump means for delivering to said analyte binding region a labelled substance which binds to analyte bound to said binding moiety to produce a detectable signal indicative of the presence of said analyte; and means for detecting the binding of the labelled substance to analyte bound to said binding moiety; said method further comprising the steps of: (iv) delivering said labelled substance to said analyte binding region to bind to said bound analyte; and (v) detecting said detectable signal with said binding means, thereby to determine the presence of the analyte.

Claim 30

Dependent Claim 13 words

The method of claim 24, wherein said detectable signal comprises a luminescent signal.

Claim 31

Dependent Claim 33 words

The method of claim 21, wherein said sample comprises a cell population, said method comprising the additional step of separating said cell population from other cells within said substrate prior to step (iii).

Claim 32

Dependent Claim 22 words

The method of claim 21, wherein the width and depth of said flow channel each are between 2.0 um and 500 um.

Claim 33

Independent Claim 199 words

The method for detecting the presence of a cell population in a sample, the method comprising the steps of: (i) providing a device comprising: a sample inlet port, said port being dimensioned for connection to a pump means; and a mesoscale flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds a surface molecule on members of said cell population, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a window disposed over said analyte binding region on said substrate; (ii) delivering a sample to said inlet port and, by a pump means, through said flow system to said binding region, to permit said binding moiety to bind to a surface molecule on members of said cell population to induce agglutination of said cell population on the surface of said binding region; and (iii) detecting said agglutination optically in said binding region through said window, thereby to determine the presence of the cell population in the sample.

Claim 34

Independent Claim 199 words

The method for detecting the presence of a cell population in a sample, the method comprising the steps of: (i) providing a device comprising: a sample inlet port, said port being dimensioned for connection to a pump means; and a mesoscale flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds a surface molecule on members of said cell population, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a window disposed over said analyte binding region on said substrate; (ii) delivering a sample to said inlet port and, by a pump means, through said flow system to said binding region, to permit said binding moiety to bind to a surface molecule on members of said cell population to induce agglutination of said cell population on the surface of said binding region; and (iii) detecting said agglutination optically in said binding region through said window, thereby to determine the presence of the cell population in the sample.

Claim 35

Independent Claim 212 words

A method for detecting the presence of one or more analytes in a fluid sample, the method comprising the steps of: (i) providing a device comprising: a solid substrate microfabricated to define: a sample inlet port; and a first and at least a second mesoscale flow system, each said flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds to analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a detection means associated with each analyte binding region or in fluid communication with each analyte binding region for detecting binding of analyte to the binding moiety in each analyte binding region, thereby to determine the presence of said analyte; (ii) delivering a sample to said inlet port and, by pump means, through said flow system to a said analyte binding regions; and (iii) detecting the binding of an analyte to the binding moiety in each of the analyte binding regions in said first and at least second system with said binding means, thereby to determine the presence of the analytes.

Claim 36

Independent Claim 212 words

A method for detecting the presence of one or more analytes in a fluid sample, the method comprising the steps of: (i) providing a device comprising: a solid substrate microfabricated to define: a sample inlet port; and a first and at least a second mesoscale flow system, each said flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising binding moiety immobilized on a surface in said analyte binding region, wherein said binding moiety specifically binds to analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; and a detection means associated with each analyte binding region or in fluid communication with each analyte binding region for detecting binding of analyte to the binding moiety in each analyte binding region, thereby to determine the presence of said analyte; (ii) delivering a sample to said inlet port and, by pump means, through said flow system to a said analyte binding regions; and (iii) detecting the binding of an analyte to the binding moiety in each of the analyte binding regions in said first and at least second system with said binding means, thereby to determine the presence of the analytes.

Claim 37

Dependent Claim 152 words

In combination, a device for determining an analyte in a fluid sample, said device comprising a solid substrate having at least one surface and being microfabricated to define: a sample inlet port; and a flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety which specifically binds to said analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; detection means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting binding of said analyte to said binding moiety; and an appliance for use with said device, said appliance comprising a holder for said device, a fluid sample input conduit interfitting with said inlet port of said device and a pump for passing fluid sample through said flow system.

Claim 38

Dependent Claim 152 words

In combination, a device for determining an analyte in a fluid sample, said device comprising a solid substrate having at least one surface and being microfabricated to define: a sample inlet port; and a flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety which specifically binds to said analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; detection means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting binding of said analyte to said binding moiety; and an appliance for use with said device, said appliance comprising a holder for said device, a fluid sample input conduit interfitting with said inlet port of said device and a pump for passing fluid sample through said flow system.

Claim 39

Dependent Claim 17 words

The combination of claim 30, wherein the analyte binding region of said device has mesoscale cross-sectional dimensions.

Claim 40

Dependent Claim 16 words

The combination of claim 30, wherein the flow channel of said device has mesoscale cross-sectional dimensions.

Claim 41

Dependent Claim 22 words

The combination of claim 30, wherein said binding moiety is immobilized on a surface within the analyte binding region of said device.

Claim 42

Dependent Claim 26 words

The combination of claim 30, wherein said appliance further comprises a fluid reservoir and means for delivering fluid contained in said reservoir to said flow system.

Claim 43

Dependent Claim 24 words

The combination as claimed in claim 34, wherein said reservoir contains a labelled reagent which produces a detectable signal when bound to said analyte.

Claim 44

Dependent Claim 31 words

The combination of claim 30, wherein said analyte binding region comprises particles having analyte binding moieties immobilized on the surface thereof which in the presence of said analyte, induce particle agglomeration.

Claim 45

Dependent Claim 31 words

The combination of claim 30, wherein the flow system of said device is microfabricated in a surface of said solid substrate and enclosed by a transparent cover adhered to said surface.

Claim 46

Dependent Claim 24 words

The combination of claim 30, wherein said analyte is a ligand and said binding moiety is a receptor which binds specifically to said ligand.

Claim 47

Dependent Claim 26 words

The combination of claim 30, wherein said analyte is an antigen and said binding moiety is an antigen binding protein which binds specifically to said antigen.

Claim 48

Dependent Claim 25 words

The combination of claim 30, wherein said analyte is a polynucleotide and said binding moiety is a complementary polynucleotide that hybridizes to said polynucleotide analyte.

Claim 49

Independent Claim 196 words

A method for determining the presence or amount of an intercellular component in a cell-containing biological fluid sample, said method comprising the steps of: (i) providing a device comprising a solid substrate microfabricated to define: a sample inlet port; and a flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety which specifically binds said analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; cell lysing means in said mesoscale flow system, and upstream of said analyte binding region, in fluid communication with said flow channel; binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting binding of said analyte to said binding moiety; (ii) delivering said sample to said inlet port; (iii) lysing said cells within said device to release said intercellular component; (iv) delivering said intercellular component to said analyte binding region; and (v) detecting binding of said intercellular component to said binding moiety as determinative of the presence or amount of said intercellular component in said sample.

Claim 50

Independent Claim 196 words

A method for determining the presence or amount of an intercellular component in a cell-containing biological fluid sample, said method comprising the steps of: (i) providing a device comprising a solid substrate microfabricated to define: a sample inlet port; and a flow system comprising: a sample flow channel extending from said inlet port; and an analyte binding region in fluid communication with said flow channel comprising a binding moiety which specifically binds said analyte, at least one of said sample flow channel or said analyte binding region having mesoscale cross-sectional dimensions; cell lysing means in said mesoscale flow system, and upstream of said analyte binding region, in fluid communication with said flow channel; binding means associated with said analyte binding region or in fluid communication with said analyte binding region for detecting binding of said analyte to said binding moiety; (ii) delivering said sample to said inlet port; (iii) lysing said cells within said device to release said intercellular component; (iv) delivering said intercellular component to said analyte binding region; and (v) detecting binding of said intercellular component to said binding moiety as determinative of the presence or amount of said intercellular component in said sample.

Claim 51

Dependent Claim 48 words

The method of claim 41, wherein said intercellular component is in a subpopulation of cells within a mixed cell population in said sample, and said method includes the additional step of separating said cell subpopulation from said mixed cell population within said device prior to said lysing step.

Claim 52

Dependent Claim 71 words

The method of claim 41, wherein the device provided in step (i) further includes means for delivering to said bound analyte binding region a labelled reagent which binds to said bound intercellular component to produce a detectable signal indicative of the binding of said intercellular component to said binding moiety, said method further comprising delivering said labelled reagent to said analyte binding region, and, in said detecting step, detecting said signal.

Claim 53

Dependent Claim 64 words

The method of claim 41, wherein the analyte binding region in said device provided in step (i) comprises particles having analyte binding sites immobilized on the surface thereof which, in the presence of said intercellular component, induce particle agglomeration, and, as a result of step (iv), said intercellular component binds to said particles to induce agglomeration; and, in said detecting step, detecting said agglomeration.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	2000-06-09	2000-12-09	2001-06-10	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	2004-06-09	2004-12-09	2005-06-10	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2008-06-09	2008-12-09	2009-06-10	$7700.00	Window Closed / Lapsed

Methods and apparatus for the detection of an analyte utilizing mesoscale flow systems

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