US4778755A
Expired / Lapsed
US
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Immunoassay method
Assignee: BIOWHITTRAKER Inc A CORP OF
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Filing: 1983-08-08
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Granted: 1988-10-18
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Expires: 2003-08-08
Abstract
Abstract In the determination of the quantity of enzyme present as a result of an immunoassay procedure, the enzyme containing component is combined with a substrate which in the presence of the enzyme yields a photometrically detectable enzymatic reaction product and a non-interfering, enzymatically inhibiting second product. The enzymatic reaction is inhibited by the addition of an inhibiting quantity of the same compound as the second product after the enzymatic reaction has continued for a time sufficient to produce photometrically detectable first products in concentrations which are a function of the quantity of enzyme present. The enzyme reaction is inhibited before the enzymatic reacton has proceeded to the point where the quantity of photometrically detectable first compound is primarily a function of the amount of substrate. This method yields a more stable test medium than methods using other enzyme inhibitors.
Inventors
- 👤 Yuh-Geng Tsay
- 👤 Susan M. Cain
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 01:32
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/54; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/542; G01N33/58; G01N33/581; Y; Y10; Y10S; Y10S435/00; Y10S435/963
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
105 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present.
Claim 2
Independent Claim
105 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present.
Claim 3
Independent Claim
131 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present, wherein the enzyme is an alkaline phosphatase, the substrate yields a fluorescent product in the presence of the enzyme, and the second product includes phosphate ion.
Claim 4
Independent Claim
131 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present, wherein the enzyme is an alkaline phosphatase, the substrate yields a fluorescent product in the presence of the enzyme, and the second product includes phosphate ion.
Claim 5
Dependent Claim
39 words
The method of claim 2 wherein the substrate is 4-methyl-umbelliferyl phosphate, and phosphate ion is added as a phosphate solution having a pH of from 8.15 to 10.10 and containing sufficient phosphate ion to substantially terminate further enzymatic reaction.
Claim 6
Independent Claim
130 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present, wherein the enzyme is an alkaline phosphatase, the substrate yields a chromophor product in the presence of enzyme, and the second product includes phosphate ion.
Claim 7
Independent Claim
130 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present, wherein the enzyme is an alkaline phosphatase, the substrate yields a chromophor product in the presence of enzyme, and the second product includes phosphate ion.
Claim 8
Dependent Claim
39 words
The method of claim 4 wherein the substrate is p-nitrophenyl phosphate, and phosphate ion is added as a phosphate solution having a pH of from 8.5 to 10.0 and containing sufficient phosphate ion to substantially terminate further enzymatic reaction.
Claim 9
Independent Claim
129 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present, wherein the enzyme is a beta-D-galactosidase, the substrate yields a fluorescent product in the presence of the enzyme, and the second product includes D-galactose.
Claim 10
Independent Claim
129 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present, wherein the enzyme is a beta-D-galactosidase, the substrate yields a fluorescent product in the presence of the enzyme, and the second product includes D-galactose.
Claim 11
Dependent Claim
26 words
The method of claim 6 wherein the substrate is 4-methylumbelliferyl-beta-D-galactoside, and a solution of D-galactose is added in quantity sufficient to substantially terminate further enzymatic action.
Claim 12
Independent Claim
129 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present, wherein the enzyme is a glucose-6-P-dehydrogenase, the substrate yields a fluorescent product in the presence of the enzyme, and the second product includes D-glucon-delta-lactone-6-phosphate.
Claim 13
Independent Claim
129 words
A method for measuring a quantity of an enzyme present in a system comprising (a) bringing together, in an aqueous medium, the enzyme and a substrate, which in the presence of the enzyme is converted to a photometrically detectable first compound and a second product for a time sufficient to produce a quantity of the first compound which is a function of the amount of enzyme present, (b) then adding additional second product to the aqueous medium in an amount sufficient to substantially terminate further enzymatic reaction, and (c) determining the quantity of the first compound as a function of the amount of enzyme present, wherein the enzyme is a glucose-6-P-dehydrogenase, the substrate yields a fluorescent product in the presence of the enzyme, and the second product includes D-glucon-delta-lactone-6-phosphate.
Claim 14
Dependent Claim
27 words
The method of claim 8 wherein the substrate is glucose-6-P and a solution of D-glucon-delta-lactone-6-phosphate is added in a quantity sufficient to substantially terminate further enzymatic action.
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| 3.5 Years (Year 4) | 1991-10-18 | 1992-04-18 | 1992-10-18 | $2000.00 | Window Closed / Lapsed |
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