EP0119623A2
Expired / Lapsed
EP
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Multilayer analytical element for non-isotopic assay
Assignee: Fujifilm Holdings Corp
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Filing: 1984-03-19
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Granted: —
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Expires: 2004-03-19
Abstract
Abstract A multilayer analytical element for non-isotopic assay of an analyte contained in an aqueous liquid sample, utilizing a competitive binding reaction to a protein between the analyte and a non-isotopically labeled analyte substance or analogue thereof, which comprises: a light-transmissive water-impermeable support; a sheet provided on the support, which comprises a continuous binder matrix containing the non-isotopically labeled analyte substance or analogue thereof; a porous reaction sheet provided on the binder matrix sheet, which contains a predetermined amount of the protein fixed therein, the protein being specifically bindable to the analyte; and a porous sharing sheet provided on the reaction sheet, in which the substantial separation of the bound non-isotopically labeled complex (B) from the free non-isotopically labeled analyte substance or analogue thereof (F) is accomplished by the presence of said porous sharing sheet. A slide enclosing the multilayer analytical element and a non-isotopic assay employing the multilayer analytical element are also disclosed.
Inventors
- 👤 Yukio Yasuda
- 👤 Nobuhito Masuda
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-25 23:41
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/74; G01N33/78; G01N33/52; G01N33/525; G01N33/53; G01N33/543; G01N33/54366; G01N33/54386; G01N33/54387; G01N33/54391; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S436/00; Y10S436/807; Y10S436/81
Claims Analysis
Total Claims: 28
Claim 1
Independent Claim
201 words
A multilayer analytical element for non-isotopic assay of an analyte contained in an aqueous liquid sample, the assay comprising a stage of subjecting the analyte, a predetermined amount of a protein specifically bindable to the analyte and a predetermined amount of a non-isotopically labeled analyte substance or analogue thereof, to competitive binding reaction; a stage of substantially separating the resulting bound non-isotopically labeled complex (B) of the protein from the free non-isotopically labeled analyte substance or analogue thereof (F); and a stage of determining the signal strength of said non-isotopically labeled complex (B), which comprises: a light-transmissive water-impermeable support; a sheet provided on the support, which comprises a continuous binder matrix containing the non-isotopically labeled analyte substance or analogue thereof in a substantially uniform amount per unit area; a porous reaction sheet provided on the continuous binder matrix sheet, which contains a predetermined amount of the protein fixed therein, the protein being specifically bindable to the analyte; and a porous sharing sheet provided on the porous reaction sheet, in which the substantial separation of the bound non-isotopically labeled complex (B) from the free non-isotopically labeled analyte substance or analogue thereof (F) is accomplished by the presence of said porous sharing sheet.
Claim 2
Dependent Claim
201 words
A multilayer analytical element for non-isotopic assay of an analyte contained in an aqueous liquid sample, the assay comprising a stage of subjecting the analyte, a predetermined amount of a protein specifically bindable to the analyte and a predetermined amount of a non-isotopically labeled analyte substance or analogue thereof, to competitive binding reaction; a stage of substantially separating the resulting bound non-isotopically labeled complex (B) of the protein from the free non-isotopically labeled analyte substance or analogue thereof (F); and a stage of determining the signal strength of said non-isotopically labeled complex (B), which comprises: a light-transmissive water-impermeable support; a sheet provided on the support, which comprises a continuous binder matrix containing the non-isotopically labeled analyte substance or analogue thereof in a substantially uniform amount per unit area; a porous reaction sheet provided on the continuous binder matrix sheet, which contains a predetermined amount of the protein fixed therein, the protein being specifically bindable to the analyte; and a porous sharing sheet provided on the porous reaction sheet, in which the substantial separation of the bound non-isotopically labeled complex (B) from the free non-isotopically labeled analyte substance or analogue thereof (F) is accomplished by the presence of said porous sharing sheet.
Claim 3
Dependent Claim
27 words
The multilayer analytical element as claimed in claim 1, in which said non-isotopically labeled analyte substance or analogue thereof is a fluorescent substance-labeled analyte substance or analogue.
Claim 4
Dependent Claim
44 words
The multilayer analytical element as claimed in claim 1, in which said non-isotopically labeled analyte substance or analogue thereof is an analyte substance or analogue thereof labeled with an enzyme, the enzymatic activity of which is capable of being modulated by an antigen-antibody reaction.
Claim 5
Dependent Claim
35 words
The multilayer analytical element as claimed in claim 1, in which said non-isotopically labeled analyte substance or analogue thereof is an analyte substance or analogue thereof having a group capable of nodulatin g a chemiluminescence.
Claim 6
Dependent Claim
51 words
The multilayer analytical element as claimed in any one of claims 1 through 4, in which: said analyte and protein are an antigen and antibody, respectively; said non-isotopically labeled analyte substance or analogue thereof is a non-isotopically labeled antigen or analogue thereof, respectively; and the binding reaction is an antigen-antibody reaction.
Claim 7
Dependent Claim
63 words
The multilayer analytical element as claimed in any claim of claims 1 through 4, in which said substantial separation of the bound non-isotopically labeled complex (B) from the free non-isotopically labeled analyte substance or analogue thereof (F) is accomplished through sharing the free non-isotopically labeled analyte substance or analogue thereof (F) between the porous reaction sheet and other space than the reaction sheet.
Claim 8
Dependent Claim
48 words
The multilayer analytical element as claimed in claim 6, in which said sharing is accomplished by adjusting the ratio of the liquid-holding capacity between the the porous sharing sheet and the porous reaction sheet to a value within the range of 1 : 3 to 5 : 1.
Claim 9
Dependent Claim
55 words
The multilayer analytical element as claimed in claim 2, in which said assay comprises determination of a total value of the fluorescence strength of the bound non-isotopically labeled complex (B) located in the porous reaction sheet and the fluorescence strength of the free non-isotopically labeled analyte substance or analogue thereof (F) located upon being shared.
Claim 10
Dependent Claim
65 words
The multilayer analytical element as claimed in claim 3, in which said assay comprises determination of strength of signals substantially provided in the presence of a substrate of the enzyme by the enzyme-labeled complex (B) located in the porous reaction sheet and by the free enzyme-labeled analyte substance or analogue thereof (F) located upon being shared, the determination being performed photometrically from the support side.
Claim 11
Dependent Claim
26 words
The multilayer analytical element as claimed in claim 9, in which said determination is performed by measuring optical absorption of a product of the enzymatic reaction.
Claim 12
Dependent Claim
29 words
The multilayer analytical element as claimed in claim 9, in which said determination is performed by measuring fluorescence strength or chemiluminescence strength of a product of the enzymatic reaction.
Claim 13
Dependent Claim
26 words
The multilayer analytical element as claimed in any claim of claims 1 through 4, in which said porous sharing sheet is made of fibrous porous material.
Claim 14
Dependent Claim
29 words
The multilayer analytical element as claimed in claim 12, in which said porous sharing sheet contains fine particles, each of which is independently distributed within the fibrous porous material.
Claim 15
Dependent Claim
20 words
The multilayer analytical element as claimed in claim 12, in which said porous sharing sheet contains a water-insoluble light-shielding material.
Claim 16
Dependent Claim
31 words
The multilayer analytical element as claimed in claim 14, in which said light-shielding material is capable of absorbing or reflecting at least a portion of light applied thereto for the determination.
Claim 17
Dependent Claim
26 words
The multilayer analytical element as claimed in any claim of claims 1 through 4, in which said porous reaction sheet is made of fibrous porous material.
Claim 18
Dependent Claim
29 words
The multilayer analytical element as claimed in claim 16, in which said porous reaction sheet contains fine particles, each of which is independently distributed within the fibrous porous material.
Claim 19
Dependent Claim
25 words
The multilayer analytical element as claimed in claim 16, in which said fibrous porous material has a void ratio of at least 60 volume %.
Claim 20
Dependent Claim
23 words
The multilayer analytical element as claimed in claim 17, in which said fine particles are made of water-insoluble polysaccharide or a derivative thereof.
Claim 21
Dependent Claim
34 words
The multilayer analytical element as claimed in claim 19, in which the surfaces of said fine particles are coated in part with a protein bindable to the analyte through physical adsorption or chemical bonding.
Claim 22
Dependent Claim
131 words
A slide for non-isotopic assay of an analyte contained in an aqueous liquid sample, in which a multilayer analytical element comprising: a light-transmissive water-impermeable support; a sheet provided on the support, which comprises a continuous binder matrix containing the non-isotopically labeled analyte substance or analogue thereof in a substantially uniform amount per unit area; a porous reaction sheet provided on the continuous binder matrix sheet, which contains a predetermined amount of the protein fixed therein, the protein being specifically bindable to the analyte; and a porous sharing sheet provided on the porous reaction sheet, is provided in a frame having on a surface of the porous sharing sheet side an opening for introduction of the liquid sample and on a surface of the support side an opening for enabling photometric measurement.
Claim 23
Dependent Claim
131 words
A slide for non-isotopic assay of an analyte contained in an aqueous liquid sample, in which a multilayer analytical element comprising: a light-transmissive water-impermeable support; a sheet provided on the support, which comprises a continuous binder matrix containing the non-isotopically labeled analyte substance or analogue thereof in a substantially uniform amount per unit area; a porous reaction sheet provided on the continuous binder matrix sheet, which contains a predetermined amount of the protein fixed therein, the protein being specifically bindable to the analyte; and a porous sharing sheet provided on the porous reaction sheet, is provided in a frame having on a surface of the porous sharing sheet side an opening for introduction of the liquid sample and on a surface of the support side an opening for enabling photometric measurement.
Claim 24
Dependent Claim
46 words
The slide as claimed in claim 21, in which said frame comprises a upper member having an opening for introduction of the liquid sample and a lower member having an opening for enabling photometric measurement, both being united through an adhesive material or direct fusion therebetween.
Claim 25
Dependent Claim
27 words
The slide as claimed in claim 22, in which at least the lower member contains a coloring material which does not substantially reflect a light for measurement.
Claim 26
Independent Claim
227 words
A method for non-isotopic assay of an analyte contained in an aqueous liquid sample comprising a stage of subjecting the analyte, a predetermined amount of a protein specifically bindable to the analyte and a predetermined amount of a non-isotopically labeled analyte substance or analogue thereof, to competitive binding reaction; a stage of substantially separating the resulting bound non-isotopically labeled complex (B) of the protein from the free non-isotopically labeled analyte substance or analogue thereof (F); and a stage of determining the signal strength of said non-isotopically labeled complex (B), which is characterized by employing a multilayer analytical element comprising: a light-transmissive water-impermeable support; a sheet provided on the support, which comprises a continuous binder matrix containing the non-isotopically labeled analyte substance or analogue thereof in a substantially uniform amount per unit area; a porous reaction sheet provided on the continuous binder matrix sheet, which contains a predetermined amount of the protein fixed therein, the protein being specifically bindable to the analyte; and a porous sharing sheet provided on the porous reaction sheet, in which the substantial separation of the bound non-isotopically labeled complex (B) from the free non-isotopically labeled analyte substance or analogue thereof (F) is accomplished by transfer of the free non-isotopically labeled analyte substance or analogue thereof (F) into a space different from the space of the porous reaction sheet upon the competitive binding reaction.
Claim 27
Independent Claim
227 words
A method for non-isotopic assay of an analyte contained in an aqueous liquid sample comprising a stage of subjecting the analyte, a predetermined amount of a protein specifically bindable to the analyte and a predetermined amount of a non-isotopically labeled analyte substance or analogue thereof, to competitive binding reaction; a stage of substantially separating the resulting bound non-isotopically labeled complex (B) of the protein from the free non-isotopically labeled analyte substance or analogue thereof (F); and a stage of determining the signal strength of said non-isotopically labeled complex (B), which is characterized by employing a multilayer analytical element comprising: a light-transmissive water-impermeable support; a sheet provided on the support, which comprises a continuous binder matrix containing the non-isotopically labeled analyte substance or analogue thereof in a substantially uniform amount per unit area; a porous reaction sheet provided on the continuous binder matrix sheet, which contains a predetermined amount of the protein fixed therein, the protein being specifically bindable to the analyte; and a porous sharing sheet provided on the porous reaction sheet, in which the substantial separation of the bound non-isotopically labeled complex (B) from the free non-isotopically labeled analyte substance or analogue thereof (F) is accomplished by transfer of the free non-isotopically labeled analyte substance or analogue thereof (F) into a space different from the space of the porous reaction sheet upon the competitive binding reaction.
Claim 28
Dependent Claim
19 words
The process for non-isotopic assay as claimed in claim 24, in which the non-isotopic assay is a non-isotopic immunoassay.
References Cited (Backward Citations)
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