US5486452A — Devices and kits for immunological analysis

Claim 1

Independent Claim 91 words

An immunological analysis device consisting of a porous sheet of nitrocellulose containing an array of preselected geometry of delimited absorption areas of at least one compound capable of specifically binding an antigen (antigen-reactive compound) which adheres tightly and does not spread out on the surface of said porous sheet, said array of preselected geometry being obtained in said porous sheet by applying liquid aliquots of said antigen-reactive compound to said porous sheet mechanically via direct contact, wherein residual absorption sited in said porous sheet are unsaturated or saturated by non-specific protein.

Claim 2

Dependent Claim 91 words

An immunological analysis device consisting of a porous sheet of nitrocellulose containing an array of preselected geometry of delimited absorption areas of at least one compound capable of specifically binding an antigen (antigen-reactive compound) which adheres tightly and does not spread out on the surface of said porous sheet, said array of preselected geometry being obtained in said porous sheet by applying liquid aliquots of said antigen-reactive compound to said porous sheet mechanically via direct contact, wherein residual absorption sited in said porous sheet are unsaturated or saturated by non-specific protein.

Claim 3

Dependent Claim 30 words

The device of claim 1 wherein said antigen-reactive compound is an immunoglobulin of human or animal origin or a binding fragment thereof for the detection and quantitation of rheumatoid factor.

Claim 4

Dependent Claim 31 words

The device of claim 1 wherein said antigen-reactive compound is a complement protein for the detection and quantitation of a circulating immune complex which circulating immune complex comprises an antigen-antibody complex.

Claim 5

Dependent Claim 18 words

The device of claim 1 wherein said porous sheet is from about 0.01 to about 0.5 mm thick.

Claim 6

Dependent Claim 14 words

The device of claim 4 wherein said porous sheet is about 0.1 mm thick.

Claim 7

Dependent Claim 18 words

The device of claim 1 wherein said nitrocellulose has about 3 nitric acid groups per 6 carbon atoms.

Claim 8

Dependent Claim 18 words

The device of claim 1 wherein said porous sheet has an average pore size of about 0.45 um.

Claim 9

Dependent Claim 13 words

The device of claim 1 wherein said array is an array of dots.

Claim 10

Dependent Claim 19 words

The device of claim 1 wherein said array is an array of microdots having diameters less than 2 mm.

Claim 11

Dependent Claim 20 words

The device of claim 1 wherein said array is an array of lines of width of 2 mm or less.

Claim 12

Dependent Claim 22 words

The device of claim 1 wherein said antigen-reactive compound is applied to said support by a capillary tube, pipette, syringe, or spray.

Claim 13

Dependent Claim 21 words

The device of claim 1 wherein said antigen-reactive compound is applied to said support in volumes of less than 1 ul.

Claim 14

Dependent Claim 31 words

The device of claim 1 wherein said residual adsorption area is saturated with a serum containing substance, wherein said substance is undiluted total serum or total serum diluted with sodium chloride.

Claim 15

Dependent Claim 38 words

The device of claim 1 wherein said antigen-reactive compound is reactive to an antigen, wherein said antigen is a constituent of human biopsy material, mammalian tissue, mammalian cells, body fluids, fungi, protozoa, metazoan parasites, bacteria, mycoplasma, or viruses.

Claim 16

Dependent Claim 30 words

A kit comprising the device of claim 1 and reagents of an indicator system in pre-aliquoted or dessicated form to allow detection of an antigen -- antigen reactive compound complex.

Claim 17

Dependent Claim 119 words

The kit of claim 15 wherein said indicator is selected from the group consisting of A) antibodies capable of forming antigen-antibody complexes, wherein the antibody is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving a color reaction with a suitable substrate, B) complement protein capable of binding to antigen-antibody complexes, wherein the complement protein is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving color reaction with a suitable substrate, and C) complement protein and a further specific anti-complement antibody, wherein the antibody is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving a color reaction with a suitable substrate.

Claim 18

Dependent Claim 60 words

The kit of claim 16, wherein said antigen-reactive compound is complement protein for the detection and quantitation of circulating immune complex which circulating immune complex comprises an antigen-antibody complex and wherein the indicator is a group A) antibody radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving a color reaction with a suitable substrate.

Claim 19

Dependent Claim 30 words

The kit of claim 15, wherein said antigen-reactive compound is an immunoglobulin of human or animal origin or a binding fragment thereof for the detection and quantitation of rheumatoid factor.

Claim 20

Dependent Claim 43 words

The kit of claim 15, comprising a multi-cavity plastic tray, lyophilized mixtures of indicator antibody, salts, buffers, carrier serum or protein, and pre-determined amounts of indicator enzyme chromogenie substrate, salts, buffers, and ampoules containing pre-measured volumes of liquid substrates, all in suitable packaging.

Claim 21

Dependent Claim 30 words

The kit of claim 15 wherein said reagents of said indicator system comprise a substrate, a cofaetor or prosthetic group for an indicator enzyme, or a coupled series of enzymes.

Claim 22

Dependent Claim 23 words

The kit of claim 15, wherein the reagents of said indicator system comprise a covalent adduct between an antigen and a signalling molecule.

Claim 23

Dependent Claim 195 words

A method of immunological analysis comprising (a) incubating the device of claim 1 wherein residual absorption sites in said porous sheet are saturated by non-specific protein, with a sample containing an immunoglobulin or other antigen to be detected; (b) incubating said device with a solution of an indicator selected from the group consisting of A) antibodies capable of forming antigen-antibody complexes, wherein the antibody is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving a color reaction with a suitable substrate, B) complement protein capable of binding to antigen-antibody complexes, wherein the complement protein is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving color reaction with a suitable substrate, and C) complement protein and a further specific anti-complement antibody, wherein the antibody is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving a color reaction with a suitable substrate; (c) when the antibody or complement protein of step (b) is conjugated with an enzyme, incubating said device with a solution of said suitable enzyme substrate; and (d) detecting the signal of said indicator of step (b).

Claim 24

Dependent Claim 32 words

The method of claim 22, wherein the incubation with a sample containing the immunogloblin or antigen to be detected is in the presence of non-specific proteins with regard to the immunoreactive compounds.

Claim 25

Dependent Claim 40 words

The method of claim 22, wherein the indicator is selected from the group consisting of radioactively labelled antibodies, antibodies conjugated with a fluorescent substance, and antibodies conjugated with an enzyme capable of giving a color reaction with a suitable substrate.

Claim 26

Dependent Claim 14 words

A method of claim 22, wherein the indicator is a horseradish peroxidase conjugated antibody.

Claim 27

Dependent Claim 82 words

An immunological analysis device consisting of a porous sheet of nitrocellulose containing an array of preselected geometry of delimited absorption areas of at least one antigen, which adheres tightly and does not spread out on the surface of said porous sheet, said array of preselected geometry being obtained in said porous sheet by applying liquid aliquots of said antigen to said porous sheet mechanically via direct contact, wherein residual absorption sites in said porous sheet are unsaturated or saturated by non-specific protein.

Claim 28

Dependent Claim 82 words

An immunological analysis device consisting of a porous sheet of nitrocellulose containing an array of preselected geometry of delimited absorption areas of at least one antigen, which adheres tightly and does not spread out on the surface of said porous sheet, said array of preselected geometry being obtained in said porous sheet by applying liquid aliquots of said antigen to said porous sheet mechanically via direct contact, wherein residual absorption sites in said porous sheet are unsaturated or saturated by non-specific protein.

Claim 29

Dependent Claim 18 words

The device of claim 26 wherein said porous sheet is from about 0.01 to about 0.5 mm thick.

Claim 30

Dependent Claim 14 words

The device of claim 27 wherein said porous sheet is about 0.1 mm thick.

Claim 31

Dependent Claim 18 words

The device of claim 27 wherein said nitrocellulose has about 3 nitric acid groups per 6 carbon atoms.

Claim 32

Dependent Claim 18 words

The device of claim 27 wherein said porous sheet has an average pore size of about 0.45 um.

Claim 33

Dependent Claim 13 words

The device of claim 27 wherein said array is an array of dots.

Claim 34

Dependent Claim 19 words

The device of claim 27 wherein said array is an array of microdots having diameters less than 2 mm.

Claim 35

Dependent Claim 20 words

The device of claim 27 wherein said array is an array of lines of width of 2 mm or less.

Claim 36

Dependent Claim 21 words

The device of claim 26 wherein said antigen is applied to said support by a capillary tube, pipette, syringe, or spray.

Claim 37

Dependent Claim 20 words

The device of claim 26 wherein said antigen is applied to said support in volumes of less than 1 ul.

Claim 38

Dependent Claim 31 words

The device of claim 26 wherein said residual adsorption area is saturated with a serum containing substance, wherein said substance is undiluted total serum a total serum diluted with sodium chloride.

Claim 39

Dependent Claim 29 words

The device of claim 26 wherein said antigen is a constituent of human biopsy material, mammalian tissue, mammalian cells, body fluids, fungi, protozoa, metazoan parasites, bacteria, mycoplasma, or viruses.

Claim 40

Dependent Claim 30 words

A kit comprising the device of claim 26 and reagents of an indicator system in pre-aliquoted or dessicated form to allow detection of an antigen -- antigen reactive compound complex.

Claim 41

Dependent Claim 118 words

The kit of claim 38 wherein said indicator is selected from the group consisting A) antibodies capable of forming antigen-antibody complexes, wherein the antibody is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving a color reaction with a suitable substrate, B) complement protein capable of binding to antigen-antibody complexes, wherein the complement protein is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving color reaction with a suitable substrate, and C) complement protein and a further specific anti-complement antibody, wherein the antibody is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving a color reaction with a suitable substrate.

Claim 42

Dependent Claim 43 words

The kit of claim 38, comprising a multi-cavity plastic tray, lyophilized mixtures of indicator antibody, salts, buffers, carrier serum or protein, and pre-determined amounts of indicator enzyme chromogenic substrate, salts, buffers, and ampoules containing pre-measured volumes of liquid substrates, all in suitable packaging.

Claim 43

Dependent Claim 30 words

The kit of claim 38 wherein said reagents of said indicator system comprise a substrate, a cofactor or prosthetic group for an indicator enzyme, or a coupled series of enzymes.

Claim 44

Dependent Claim 23 words

The kit of claim 38, wherein the reagents of said indicator system comprise a covalent adduct between an antigen and a signalling molecule.

Claim 45

Dependent Claim 192 words

A method of immunological analysis comprising (a) incubating the device of claim 26 wherein residual absorption sites in said porous sheet are saturated by non-specific protein, with a sample containing an immunoglobulin to be detected; (b) incubating said device with a solution of an indicator selected from the group consisting of A) antibodies capable of forming antigen-antibody complexes, wherein the antibody is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving a color reaction with a suitable substrate, B) complement protein capable of binding to antigen-antibody complexes, wherein the complement protein is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving color reaction with a suitable substrate, and C) complement protein and a further specific anti-complement antibody, wherein the antibody is radioactively labelled, conjugated with a fluorescent substance or conjugated with an enzyme capable of giving a color reaction with a suitable substrate; (c) when the antibody or complement protein of step (b) is conjugated with an enzyme, incubating said device with a solution of said suitable enzyme substrate; and (d) detecting the signal of said indicator of step (b).

Claim 46

Dependent Claim 30 words

The method of claim 43, wherein the incubation with a sample containing the immunogloblin to be detected is in the presence of non-specific proteins with regard to the immunoreactive compounds.

Claim 47

Dependent Claim 40 words

The method of claim 43, wherein the indicator is selected from the group consisting of radioactively labelled antibodies, antibodies conjugated with a fluorescent substance, and antibodies conjugated with an enzyme capable of giving a color reaction with a suitable substrate.

Claim 48

Dependent Claim 14 words

A method of claim 43, wherein the indicator is a horseradish peroxidase conjugated antibody.

Claim 49

Independent Claim 141 words

A method of producing an immunoassay device which comprises the steps of: (a) applying droplets of substantially enriched aqueous solutions of antibodies of distinct specification to a substantially planar surface of a support, locating said droplets so that they cover small, discrete, closely spaced areas of the planar surface of the support and form an array; (b) allowing said aqueous solutions of antibodies to remain on said areas of the surface for a period of time long enough for the antibodies in solution to be adsorbed to said areas of the surface to form discrete, antibody-coated areas; (c) swiftly flooding the entire surface of the solid support with an aqueous solution of protein in order to block non-specific binding sites in areas of the surface of the solid support which surround the antibody-coated areas; and (d) washing and drying the device.

Claim 50

Independent Claim 141 words

A method of producing an immunoassay device which comprises the steps of: (a) applying droplets of substantially enriched aqueous solutions of antibodies of distinct specification to a substantially planar surface of a support, locating said droplets so that they cover small, discrete, closely spaced areas of the planar surface of the support and form an array; (b) allowing said aqueous solutions of antibodies to remain on said areas of the surface for a period of time long enough for the antibodies in solution to be adsorbed to said areas of the surface to form discrete, antibody-coated areas; (c) swiftly flooding the entire surface of the solid support with an aqueous solution of protein in order to block non-specific binding sites in areas of the surface of the solid support which surround the antibody-coated areas; and (d) washing and drying the device.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1999-01-22	1999-07-24	2000-01-23	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	2003-01-22	2003-07-24	2004-01-23	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2007-01-22	2007-07-24	2008-01-23	$7700.00	Window Closed / Lapsed

Devices and kits for immunological analysis

Abstract

Inventors

Bibliographic Details

Claims Analysis

References Cited (Backward Citations)

USPTO Annuity & Maintenance Windows

Assigned Categories & Tags

Assign Category Tag