EP0271204A2 — Immunochromatographic method and device

Claim 1

Independent Claim 190 words

A method for determining the presence of an analyte in a sample suspected of containing said analyte, which analyte is a member of a specific binding pair ("shop member") consisting of ligand and its complementary receptor, which method comprises - (a) contacting, with a test solution containing said sample and a first sbp member analogous to said analyte, a contact portion of a piece of bibulous material capable of being traversed in at least one direction by said test solution by capillary migration, said bibulous material containing a second sbp member capable of binding said analyte and said first sbp member non-diffusively bound thereto at least at a portion thereof between said contact portion and a small situs on said bibulous material separated from said contact portion, the surface area of said situs being substantially less than that of said piece of bibulous material, said situs being capable of binding said first sbp member, (b) allowing at least a portion of the test solution to traverse said piece of bibulous material by means of capillary migration and thereby contact said situs, (c) detecting said first sbp member at said situs.

Claim 2

Independent Claim 190 words

A method for determining the presence of an analyte in a sample suspected of containing said analyte, which analyte is a member of a specific binding pair ("shop member") consisting of ligand and its complementary receptor, which method comprises - (a) contacting, with a test solution containing said sample and a first sbp member analogous to said analyte, a contact portion of a piece of bibulous material capable of being traversed in at least one direction by said test solution by capillary migration, said bibulous material containing a second sbp member capable of binding said analyte and said first sbp member non-diffusively bound thereto at least at a portion thereof between said contact portion and a small situs on said bibulous material separated from said contact portion, the surface area of said situs being substantially less than that of said piece of bibulous material, said situs being capable of binding said first sbp member, (b) allowing at least a portion of the test solution to traverse said piece of bibulous material by means of capillary migration and thereby contact said situs, (c) detecting said first sbp member at said situs.

Claim 3

Dependent Claim 36 words

The method of Claim 1 wherein said first sbp member is a conjugate of said analyte, a label, and a third sbp member and receptor for said third sbp member is non-diffusively bound to said situs.

Claim 4

Dependent Claim 31 words

The method of Claim 1 or 2 wherein a high density of said second sbp member is provided at said situs compared to portions of said strip adjacent to said situs.

Claim 5

Independent Claim 21 words

The method of any one of the preceding Claims wherein said second sbp member at said situs is bound to particles.

Claim 6

Independent Claim 19 words

The method of any one of the preceding Claims wherein said first sbp member is conjugated to a label.

Claim 7

Dependent Claim 11 words

The method of claim 5 wherein said label is an enzyme.

Claim 8

Independent Claim 33 words

The method of any one of the preceding Claims wherein the analyte is a drug, a drug analog is conjugated to a label, and said second sbp member is antibody to said drug.

Claim 9

Independent Claim 29 words

The method of any one of the preceding Claims wherein the small situs is a band transverse to the direction of traversal of said test solution along said piece.

Claim 10

Independent Claim 47 words

The method of any one of the preceding claims wherein the signal produced at the small situs has a sharp-edged distinctive pattern that provides a sharp contrast to signal produced at adjacent sites on said piece of bibulous material when analyte is present in said test solution.

Claim 11

Dependent Claim 50 words

The method of Claim 6 wherein a second enzyme is bound uniformly to said piece at least over an area including said situs and the portions of the piece adjacent to said situs, the enzymes being related in that the product of one enzyme is the substrate for the other.

Claim 12

Independent Claim 72 words

The method of any one of the preceding Claims for identifying which of a plurality of analytes are present in said test solution, wherein a plurality of second sbp members are each non-diffusively bound to said strip and each of said second sbp members is respectively capable of specifically binding a corresponding first sbp member and wherein a plurality of situses are employed, each respectively capable of binding a first sbp member.

Claim 13

Independent Claim 25 words

The method of any one of the preceding Claims wherein said test solution is allowed to traverse said piece through said situs in step (b).

Claim 14

Independent Claim 39 words

The method of any one of the preceding Claims wherein a high density of a binding agent capable of binding said first sbp member is provided at said situs compared to portions of said piece adjacent to said situs.

Claim 15

Independent Claim 28 words

The method of any one of the preceding Claims wherein said first sbp member is detected at said situs by directly detecting a detectible signal at said situs.

Claim 16

Dependent Claim 233 words

A method according to Claim 1 for determining the presence of an analyte in a sample suspected of containing said analyte, which comprises - (a) contacting, with a test solution containing said sample and an analyte analog conjugated to an enzyme, the end portion of a strip of bibulous material capable of being traversed by said test solution by capillary migration, said strip containing antibody for said analyte non--diffusively bound thereto at least at a portion thereof between a small situs on said strip separated from said end portion and said end portion, the surface area of said situs being substantially less than that of said strip, said situs having non-diffusively bound thereto a binding agent for said conjugated analyte analog at a density higher than the density of said antibody bound to said strip adjacent to said situs, (b) allowing at least a portion of the test solution to traverse said strip by capillary migration, (c) contacting said strip with a developer solution containing members of a signal producing system other than said enzyme to provide contact of said developer solution with said situs said signal producing system being capable of producing a detectible signal at said situs in relation to the amount of analyte in the test solution, and (d) comparing said detectible signal at said situs with the signal detectible at a portion of the strip adjacent to said situs.

Claim 17

Dependent Claim 48 words

The method of Claim 15 for determining a plurality of analytes in said sample wherein a specific antibody to each analyte is non-diffusively bound to said strip and wherein a plurality of situses are employed, each respectively having a receptor specific for conjugated analyte analog non-diffusively bound thereto.

Claim 18

Dependent Claim 17 words

The method of claim 15 or 16 wherein said binding agent is an antibody for an analyte.

Claim 19

Dependent Claim 27 words

The method of Claim 15, 16 or 17 wherein said small situs is a band traverse to the direction of traversal of said sample along said strip.

Claim 20

Independent Claim 45 words

The method of any one of Claims 15 to 18 wherein the signal produced at the small situs has a sharp-edged distinctive pattern that provides a sharp contrast to signal produced at adjacent sites on said strip when analyte is present in said test solution.

Claim 21

Independent Claim 129 words

a device for determining the presence of an analyte in a test solution comprising a sample suspected of containing said analyte and a first sbp member, which analyte is a member of a specific binding pair, said device comprising a bibulous strip capable of traversal by said test solution by capillary migration, said strip having a contact portion for contacting said test solution and a second sbp member complementary to said analyte non-diffusively bound to said strip at least at a portion thereof between said contact portion and a small situs on said strip separated from said contact portion, the surface area of said situs being substantially less than that of said strip, said situs having non-diffusively bound thereto a binding agent capable of binding said first sbp member.

Claim 22

Independent Claim 129 words

a device for determining the presence of an analyte in a test solution comprising a sample suspected of containing said analyte and a first sbp member, which analyte is a member of a specific binding pair, said device comprising a bibulous strip capable of traversal by said test solution by capillary migration, said strip having a contact portion for contacting said test solution and a second sbp member complementary to said analyte non-diffusively bound to said strip at least at a portion thereof between said contact portion and a small situs on said strip separated from said contact portion, the surface area of said situs being substantially less than that of said strip, said situs having non-diffusively bound thereto a binding agent capable of binding said first sbp member.

Claim 23

Dependent Claim 119 words

A device according to Claim 20 for determining the presence of an analyte in a test solution comprising a sample suspected of containing said analyte and an analyte analog, said device comprising a bibulous strip capable of traversal by said test solution by capillary migration, said strip having a contact portion for contacting said test solution and an antibody for said analyte non-diffusively bound to said strip at least at a portion thereof between said contact portion and a small situs on said strip separated from said contact portion, the surface area of said situs being substantially less than that of said strip, said situs having a non-diffusively bound thereto a binding agent capable of binding said analyte analog.

Claim 24

Dependent Claim 42 words

The device of Claim 21 for determining the presence of a plurality of analytes in said test solution wherein each of a set of antibodies is respectively non-diffusively bound to said strip and to each of different small situses on said strip.

Claim 25

Dependent Claim 66 words

A kit for use in determining the presence of an analyte in a test solution, said kit comprising in a packaged combination-- (a) the device of Claim 20 (b) said first sbp member as part of a signal producing system, (c) other members of signal producing system for producing a detectible signal in the presence of said analyte as required, and (d) ancillary materials as required.

Claim 26

Dependent Claim 230 words

A method according to Claim 1 for determining the presence of one or more of a plurality of analytes in a sample suspected of containing said analytes, each analyte being a member of a specific binding pair ("sbp member") consisting of ligand and its complementary receptor, which method comprises - (a) contacting, with a test solution containing said sample and a plurality of first sbp members each respectively analogous to one of said analytes, a contact portion of a piece of bibulous material capable of being traversed in at least one direction by said test solution by capillary migration, said bibulous material containing a plurality of second sbp members each respectively capable of binding one of said analytes and said first sbp members non-diffusively bound thereto at a portion thereof at least between said contact portion and one or more of a plurality of small situses on said bubulous material separated from said contact portion, the surface area of each situs being substantially less than that of said piece of bibulous material, each situs being capable of binding at least one of said first sbp members. (b) allowing at least a portion of the test solution to traverse said piece of bibulous material by means of capillary migration and thereby contact said situs, (c) detecting one or more of said first sbp members at one or more of said situses.

Claim 27

Dependent Claim 37 words

The method of claim 24 wherein said first sbp member is a conjugate comprising one of said analytes, an enzyme, and a third sbp member and said binding agent is a receptor for said third sbp member.

Claim 28

Dependent Claim 70 words

The method of Claim 24 wherein each of said first sbp members is conjugated to a label, the label is an enzyme and a second enzyme is bound uniformly to said piece at least over an area including said situses and the portions of the piece of bibulous material adjacent to said situses, the enzymes being related in that the product of one enzyme is the substrate for the other.

Claim 29

Dependent Claim 146 words

A device according to Claim 20 for determining the presence of one or more of a plurality of analytes in a test solution, each analyte being a member of a specific binding pair, said device comprising a piece of bibulous material capable of traversal in at least one direction by said test solution by capillary migration, said bibulous material having a contact portion for contacting said test solution and one or more of a plurality of second sbp members each complementary to one of said analytes non-diffusively bound to said bibulous material at least between said contact portion and one or more small situses on said bibulous material separated from said contact portion, the surface area of each situs being substantially less than that of said bibulous material, each situs having non-diffusively bound thereto at least one binding agent capable of binding said first sbp member.

Claim 30

Dependent Claim 15 words

The device of Claim 27 wherein said piece of bibulous material is constructed in segments.

Claim 31

Dependent Claim 70 words

A kit according to Claim 23 for use in determining the presence of an analyte in a test solution, said kit comprising in a packaged combination-- (a) the device of Claim 27 (b) said first sbp member as part of a signal producing system, (c) other members of signal producing system for producing a detectible signal in the presence of said analyte as required, and (d) ancillary materials as required.

Claim 32

Dependent Claim 255 words

A method according to Claim 1 for determining the presence of one or more of a plurality of analytes, each analyte being a member of a specific binding pair (sbp) consisting of ligand and its complementary receptor, in a sample suspected of containing one or more of said analytes, which comprises - (a) contacting, with a test solution containing said sample and one or more of a plurality of conjugates each comprising one of said analytes, a label, and an sbp member, a contact portion of a piece comprised of bibulous material capable of being traversed in at least one direction by said test solution by capillary migration, said bibulous material containing non-diffusively bound to a portion thereof at least between said contact portion and one or more situses separate from said contact portion a plurality of first receptors each respectively capable of binding to one of said analytes, said bibulous material further containing non-diffusively bound thereto at one or more situses one or more of a plurality of second receptors each respectively capable of binding to a corresponding sbp member of said conjugate, the surface area of said situs being less than that of said bibulous material, said situs being separated from said portion of said bibulous material containing one or more first receptors, (b) allowing at least a portion of said test solution to traverse said bibulous material by capillary migration and thereby contact one or more situses, and (c) examining said situses for the presence of one or more of said conjugates.

Claim 33

Dependent Claim 41 words

The method of Claim 30 wherein said analytes are morphine and tetrahydrocannabinol, said conjugates are morphine-horseradish peroxdase-fluorescein and tetrahydrocannabinol-horseradish peroxidase-biotin, said first receptors are antibodies for morphine and antibodies for tetrahydrocannabinol, and said second receptors are antibodies for fluorescein and avidin.

Claim 34

Dependent Claim 196 words

A device according to Claim 20 for determining the presence of one or more of a plurality of analytes, each analyte being a member of a specific binding pair (sbp) consisting of ligand and complementary receptor, in a test solution comprised of (i) one or more conjugates comprising one of said analytes, a label, and an sbp member and (ii) a sample suspected of containing the analyte, said device comprising - a piece comprised of bibulous material capable of traversal in at least one direction by said test solution by capillary migration, said bibulous material having a contact portion for contacting said test solution and one or more of a plurality of first receptors each respectively capable of binding to one of said analytes non-diffusively bound to said bibulous material at a portion lying at least between said contact portion and one or more situses, the surface area of each situs being less than that of said bibulous material, each situs having non-diffusively bound thereto one or more of a plurality of second receptors each respectively capable of binding an sbp member, said situs being separated from said portion of said strip containing said first receptor.

Claim 35

Dependent Claim 63 words

A kit according to Claim 23 for use in determining the presence of one or more of a plurality of analytes in a test solution, comprising in a packaged combination - (a) one or more of a plurality of conjugates each comprising one of said analytes, a label, and a member of a specific binding pair, and (b) the device of Claim 32.

Claim 36

Independent Claim 69 words

A device for determining the presence of an analyte in a test solution, said device comprising a piece of bibulous material containing a segment to which a member of a specific binding pair (sbp) complementary to said analyte is homogeneously and non-diffusively bound, characterised by having a situs on said bibulous material, said situs having non-diffusively bound thereto a binding agent capable of binding an analog of said analyte.

Claim 37

Independent Claim 69 words

A device for determining the presence of an analyte in a test solution, said device comprising a piece of bibulous material containing a segment to which a member of a specific binding pair (sbp) complementary to said analyte is homogeneously and non-diffusively bound, characterised by having a situs on said bibulous material, said situs having non-diffusively bound thereto a binding agent capable of binding an analog of said analyte.

Claim 38

Dependent Claim 59 words

A kit for use in determining the presence of one or more of a plurality of analytes in a test solution, comprising in a packaged combination - (a) one or more of a plurality of conjugates each comprising one of said analytes, a label, and a member of a specific binding pair, and (b) the device of Claim 35.

Immunochromatographic method and device

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