US4670381A — Heterogeneous immunoassay utilizing horizontal se…

Claim 1

Independent Claim 122 words

A method for the determination of an immunologically reactive ligand in a liquid using a dry analytical element, said element comprising a support having thereon a porous spreading layer which contains an immobilized receptor for an immunologically reactive ligand, said method consisting essentially of the steps of: A. in the presence of a labeled ligand analog, contacting a finite area of said spreading layer with a sample of said liquid so as to form an immobilized ligand-receptor complex within said finite area and to effect substantially horizontal separation of uncomplexed ligand from said immobilized complex during liquid spreading, and B. at least about 5 seconds after the completion of said contacting, determining said immobilized complex within the center of said finite area.

Claim 2

Independent Claim 122 words

A method for the determination of an immunologically reactive ligand in a liquid using a dry analytical element, said element comprising a support having thereon a porous spreading layer which contains an immobilized receptor for an immunologically reactive ligand, said method consisting essentially of the steps of: A. in the presence of a labeled ligand analog, contacting a finite area of said spreading layer with a sample of said liquid so as to form an immobilized ligand-receptor complex within said finite area and to effect substantially horizontal separation of uncomplexed ligand from said immobilized complex during liquid spreading, and B. at least about 5 seconds after the completion of said contacting, determining said immobilized complex within the center of said finite area.

Claim 3

Dependent Claim 25 words

The method of claim 1 wherein said immobilized complex is determined within an about 3 to about 5 mm diameter center of said finite area.

Claim 4

Dependent Claim 26 words

The method of claim 1 wherein step B is carried out within about 5 to about 180 seconds after the completion of said contacting step A.

Claim 5

Dependent Claim 25 words

The method of claim 1 wherein said liquid sample is applied to said element in a single sample volume over at least about 5 seconds.

Claim 6

Dependent Claim 27 words

The method of claim 1 wherein said liquid sample is applied to said element in two or more sequential sample volumes over at least about 5 seconds.

Claim 7

Dependent Claim 12 words

The method of claim 1 for the determination of a therapeutic drug.

Claim 8

Dependent Claim 13 words

The method of claim 6 for the determination of theophylline, phenobarbital or diphenylhydantoin.

Claim 9

Dependent Claim 24 words

The method of claim 1 wherein said ligand analog comprises an enzyme label and said element comprises an indicator composition in a reagent layer.

Claim 10

Dependent Claim 16 words

The method of claim 8 wherein said enzyme label is glucose oxidase, peroxidase or alkaline phosphatase.

Claim 11

Dependent Claim 67 words

The method of claim 1 wherein said spreading layer is composed of a particulate structure comprising a plurality of particles having a particle size of from about 1 to about 200 μm and being bonded to each other on surface areas of adjacent particles where said adjacent particles are in closest proximity to form a coherent, three dimensional lattice which is essentially non-swellable in an aqueous liquid.

Claim 12

Dependent Claim 23 words

The method of claim 10 wherein said Particles are chemically bonded to each other through reactive groups which are incorporated in said particles.

Claim 13

Dependent Claim 17 words

The method of claim 10 wherein said particles are bonded to each other with an adhesive material.

Claim 14

Dependent Claim 360 words

The method of claim 12 wherein said particles comprise an addition polymer formed from one or more of the following ethylenically unsaturated polymerizable monomers: (a) up to 100 weight percent of an aminosubstituent-free vinyl aromatic monomer, (b) up to about 25 weight percent of an acrylic acid ester, (c) up to 100 percent of a methacrylic acid ester, (d) up to about 30 weight percent of an ethylenically unsaturated carboxylic acid, (e) up to about 75 weight percent of an ethylenically unsaturated nitrile, (f) up to about 20 weight percent of an amino-substituted vinyl aromatic monomer, (g) up to about 20 weight percent of an ethylenically unsaturated monomer containing a crosslinkable group, (h) up to about 20 weight percent of a tertiary aminoalkyl acrylate or methacrylate, (i) up to 100 weight percent of an N-heterocyclic vinyl monomer, and (j) up to about 20 weight percent of an acrylamide or methacrylamide, and said adhesive comprises an addition polymer formed from a blend of ethylenically unsaturated polymerizable monomers selected from the following group: A. a blend containing from about 1 to about 35 weight percent of one or more amino-substituent-free vinyl aromatics and from about 65 to about 99 weight percent of one or more alkyl acrylates or methacrylates, B. a blend containing from about 20 to about 95 weight percent of one or more amino-substituent-free vinyl aromatics, acrylic or methacrylic acid esters or ethylenically unsaturated polymerizable monomers containing crosslinkable groups, and from about 5 to about 80 weight percent of one or more ethylenically unsaturated polymerizable monomers having an active hydrogen or salts thereof, C. a blend containing from about 15 to 100 weight percent of one or more ethylenically unsaturated monomers selected from the group consisting of 1-vinylimidazole, N-vinyl-2-pyrrolidone, vinylbenzyl alcohol, ethyl acrylate or an acrylamide or methacrylamide, and up to 85 weight percent of one or more ethylenically unsaturated polymerizable monomers containing crosslinkable groups, and D. a blend containing from about 60 to about 98 weight percent of one or more acrylic or methacrylic acid esters, and from about 2 to about 40 weight percent of one or more ethylenically unsaturated polymerizable monomers containing one or mole anionic moieties.

Claim 15

Independent Claim 180 words

A method for the determination of an immunologically reactive ligand in a liquid using a dry analytical element, said element comprising a support having thereon a porous spreading layer containing an immobilized receptor for an immunologically reactive ligand, said layer being composed of a particulate structure comprising a plurality of particles having a particle size of from about 2 to about 20 μm and being bonded to each other on surface areas of adjacent particles where said adjacent particles are in closest proximity to form a coherent, three-dimensional lattice which is essentially nonswellable in an aqueous liquid, said method consisting essentially of the steps of: A. in the presence of a labelled ligand analog, contacting a finite area of said spreading layer with a sample of said liquid so as to form an immobilized ligand-receptor complex within said finite area, and to effect substantially horizontal separation of uncomplexed ligand from said immobilized complex during liquid spreading, and B. at least about 5 seconds after the completion of said contacting, determining said immobilized complex within the center of said finite area.

Claim 16

Independent Claim 180 words

A method for the determination of an immunologically reactive ligand in a liquid using a dry analytical element, said element comprising a support having thereon a porous spreading layer containing an immobilized receptor for an immunologically reactive ligand, said layer being composed of a particulate structure comprising a plurality of particles having a particle size of from about 2 to about 20 μm and being bonded to each other on surface areas of adjacent particles where said adjacent particles are in closest proximity to form a coherent, three-dimensional lattice which is essentially nonswellable in an aqueous liquid, said method consisting essentially of the steps of: A. in the presence of a labelled ligand analog, contacting a finite area of said spreading layer with a sample of said liquid so as to form an immobilized ligand-receptor complex within said finite area, and to effect substantially horizontal separation of uncomplexed ligand from said immobilized complex during liquid spreading, and B. at least about 5 seconds after the completion of said contacting, determining said immobilized complex within the center of said finite area.

Claim 17

Dependent Claim 24 words

The method of claim 14 wherein said ligand analog comprises an enzyme label and said element comprises an indicator composition in a reagent layer.

Claim 18

Dependent Claim 19 words

The method of claim 14 wherein said ligand analog is incorporated into said element prior to contacting step A.

Claim 19

Dependent Claim 47 words

The method of claim 14 wherein said immobilized complex is determined within an about 3 to about 5 mm diameter center of said finite area, and said step B is carried out within about 5 to about 180 seconds after the completion of said contacting step A.

Claim 20

Dependent Claim 13 words

The method of claim 14 for the determination of theophylline, phenobarbital or diphenylhydantoin.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1990-06-01	1990-12-01	1991-06-02	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1994-06-01	1994-12-01	1995-06-02	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	1998-06-01	1998-12-01	1999-06-02	$7700.00	Window Closed / Lapsed

Heterogeneous immunoassay utilizing horizontal separation in an analytical element

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