Patent US5547848A successfully refreshed from Google Patents.
US5547848A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Immunoassay element containing a pulverized water-insoluble polysaccharide and process for immunoassay
Assignee: Fujifilm Holdings Corp
•
Filing: 1995-02-23
•
Granted: 1996-08-20
•
Expires: 2015-02-23
Abstract
Abstract An immunoassay element for quantitatively analyzing a ligand by determining the change in enzymatic activity. When the ligand is a low molecular weight antigen, competitive reactions between the ligand, enzyme-labelled antibody and conjugate of the antigen and high molecular weight compound are utilized. When the ligand is a macromolecular antigen, a reaction between the ligand and an enzyme-labelled antibody is utilized directly. The immunoassay element comprises a substrate layer containing a non-diffusible substrate which forms a diffusible material in the presence of the enzyme, and a reagent layer for detecting the thus formed diffusible material. The non-diffusible substrate is composed of a pulverized insoluble polysaccharide. The reagent layer may further contain a fragmenting enzyme for further fragmenting the non-diffusible material. Also provided are processes for quantitatively analyzing both of low molecular weight and macromolecular antigens contained in any samples by the use of the immunoassay elements of the invention.
Inventors
- 👤 Hiroshi Shinoki
- 👤 Toshikage Hiraoka
- 👤 Masashi Ogawa
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 14:26
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/558
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
181 words
An immunoassaying process for quantitatively analyzing an amount of a low molecular weight antigen in a sample by determining the change in enzymatic activity caused by competitive reactions between an enzyme-labelled antibody and said low molecular weight antigen and a conjugate of said low molecular weight antigen with a high molecular weight compound, comprising the steps of: (a) mixing the sample containing the low molecular weight antigen, the enzyme-labelled antibody, and the conjugate of the antigen and the high molecular weight compound so as to allow the competitive reactions; (b) applying a reaction product mixture of the competitive reactions of step (a) on a substrate layer containing a non-diffusible substrate for forming a diffusible material in the presence of said enzyme, said non-diffusible substrate being a pulverized insoluble polysaccharide which is classified after pulverization and has an average particle diameter of from 3 to 5 μm; (c) allowing to migrate said diffusible material formed in said substrate layer into a reagent layer for detecting said diffusible material; and (d) measuring the amount of said diffusible material migrating into said reagent layer.
Claim 2
Dependent Claim
181 words
An immunoassaying process for quantitatively analyzing an amount of a low molecular weight antigen in a sample by determining the change in enzymatic activity caused by competitive reactions between an enzyme-labelled antibody and said low molecular weight antigen and a conjugate of said low molecular weight antigen with a high molecular weight compound, comprising the steps of: (a) mixing the sample containing the low molecular weight antigen, the enzyme-labelled antibody, and the conjugate of the antigen and the high molecular weight compound so as to allow the competitive reactions; (b) applying a reaction product mixture of the competitive reactions of step (a) on a substrate layer containing a non-diffusible substrate for forming a diffusible material in the presence of said enzyme, said non-diffusible substrate being a pulverized insoluble polysaccharide which is classified after pulverization and has an average particle diameter of from 3 to 5 μm; (c) allowing to migrate said diffusible material formed in said substrate layer into a reagent layer for detecting said diffusible material; and (d) measuring the amount of said diffusible material migrating into said reagent layer.
Claim 3
Dependent Claim
147 words
An immunoassaying process for quantitatively analyzing an amount of a macromolecular antigen in a sample by determining the change in enzymatic activity caused by a reaction between an enzyme-labelled antibody and said macromolecular antigen comprising the steps of: (a) mixing the sample containing the macromolecular antigen and the enzyme-labelled antibody so as to allow the reaction; (b) applying a reaction product mixture of the reaction of step (a) on a substrate layer containing a non-diffusible substrate for forming a diffusible material in the presence of said enzyme, said non-diffusible substrate being a pulverized insoluble polysaccharide which is classified after pulverization and has an average particle diameter of from 3 to 5 μm; (c) allowing to migrate said diffusible material formed in said substrate layer into a reagent layer for detecting said diffusible material; and (d) measuring the amount of said diffusible material migrating into said reagent layer.
Claim 4
Dependent Claim
147 words
An immunoassaying process for quantitatively analyzing an amount of a macromolecular antigen in a sample by determining the change in enzymatic activity caused by a reaction between an enzyme-labelled antibody and said macromolecular antigen comprising the steps of: (a) mixing the sample containing the macromolecular antigen and the enzyme-labelled antibody so as to allow the reaction; (b) applying a reaction product mixture of the reaction of step (a) on a substrate layer containing a non-diffusible substrate for forming a diffusible material in the presence of said enzyme, said non-diffusible substrate being a pulverized insoluble polysaccharide which is classified after pulverization and has an average particle diameter of from 3 to 5 μm; (c) allowing to migrate said diffusible material formed in said substrate layer into a reagent layer for detecting said diffusible material; and (d) measuring the amount of said diffusible material migrating into said reagent layer.
Claim 5
Dependent Claim
125 words
In an immunoassay element for quantitatively analyzing a low molecular weight antigen by determining the change in enzymatic activity caused by competitive reactions between an enzyme-labelled antibody and said low molecular weight antigen and a conjugate of said low molecular weight antigen with a high molecular weight compound, or a macromolecular antigen by determining the change in enzymatic activity caused by a reaction between an enzyme-labelled antibody and said macromolecular antigen, wherein said element comprises a substrate layer containing a non-diffusible substrate which forms a diffusible material in the presence of said enzyme and a reagent layer for detecting said diffusible material; the improvement which comprises said non-diffusible substrate being a pulverized water-insoluble polysaccharide having an average particle diameter of from 3 to 10 μm.
Claim 6
Dependent Claim
125 words
In an immunoassay element for quantitatively analyzing a low molecular weight antigen by determining the change in enzymatic activity caused by competitive reactions between an enzyme-labelled antibody and said low molecular weight antigen and a conjugate of said low molecular weight antigen with a high molecular weight compound, or a macromolecular antigen by determining the change in enzymatic activity caused by a reaction between an enzyme-labelled antibody and said macromolecular antigen, wherein said element comprises a substrate layer containing a non-diffusible substrate which forms a diffusible material in the presence of said enzyme and a reagent layer for detecting said diffusible material; the improvement which comprises said non-diffusible substrate being a pulverized water-insoluble polysaccharide having an average particle diameter of from 3 to 10 μm.
Claim 7
Dependent Claim
14 words
The immunoassay element of claim 3, wherein said pulverized insoluble polysaccharide is carboxymethylated starch.
Claim 8
Dependent Claim
19 words
The immunoassay element of claim 3, wherein said pulverized insoluble polysaccharide is denaturated by an alkali or thermal treatment.
Claim 9
Dependent Claim
18 words
The immunoassay element of claim 4, wherein said carboxymethylated starch is denaturated by an alkali or thermal treatment.
Claim 10
Dependent Claim
16 words
The immunoassay element of claim 7, wherein said enzyme labelling to said antigen is a glucosidase.
Claim 11
Dependent Claim
11 words
The immunoassay element of claim 3, wherein said glucosidase is α-amylase.
Claim 12
Dependent Claim
36 words
The immunoassay element of claim 3, further comprising a fragmenting enzyme for fragmenting said diffusible material into a lower molecular weight product and contained in said reagent layer or another layer laminated under said reagent layer.
Claim 13
Dependent Claim
42 words
The immunoassay element of claim 9, further comprising a reagent composition for reacting with said lower molecular weight product to form a dye having an absorption peak in the visible wavelength range and contained in said reagent layer or another water-permeable layer.
Claim 14
Dependent Claim
24 words
The immunoassay element of claim 3, wherein said enzyme-lablelled antibody is contained in said substrate layer or another layer laminated on said substrate layer.
References Cited (Backward Citations)
US4066509A
Cited
US4102747A
Cited
EP0005148A1
Cited
US4337310A
Cited
EP0034692A2
Cited
EP0134291A1
Cited
JPS6131954A
Cited
EP0347839A2
Cited
US5093081A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1999-08-20 | 2000-02-19 | 2000-08-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-08-20 | 2004-02-19 | 2004-08-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-08-20 | 2008-02-19 | 2008-08-20 | $7700.00 | Window Closed / Lapsed |