US4681841A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Enzymatic assay method
Assignee: Asahi Kasei Corp
•
Filing: 1983-10-03
•
Granted: 1987-07-21
•
Expires: 2003-10-03
Abstract
Abstract A method for assaying a compound of the formula <IMAGE> wherein R1 is hydroxyl or amino, or hydrogen if at least one of R2, R3, R4, R5 and R6 is hydroxyl or amino, and R2, R3, R4, R5 and R6 are hydrogen, halogen, lower alkyl, lower alkoxy, amino, substituted amino, hydroxy, carboxyl or sulfo, or R5 and R6 together form a ring, comprises establishing a reaction system containing the compound to be assayed and a coupler and an enzyme capable of consuming oxygen and generating a pigment in the presence of the compound and the coupler. A detectable change in the reaction system is measured, to assay the compound in question. The measurement can comprise the measurement of consumed oxygen, as by an oxygen electrode. Or the measurement can comprise the measurement of the generated pigment, as by a colorimetric assay at a specific absorption wavelength thereof.
Inventors
- 👤 Kunio Matsumoto
- 👤 Tsutomu Hirata
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 08:39
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/001; C12Q1/004; C12Q1/34; C12Q2334/00; C12Q2337/00; C12Q2337/10; G; G01; G01N; G01N2333/00; G01N2333/90; G01N2333/914; G01N2333/924; G01N2333/948; G01N2333/976; Y; Y10; Y10S; Y10S435/00; Y10S435/817
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
143 words
A method for assaying a compound of the formula ##STR10## wherein R 1 is hydroxyl or amino, or hydrogen if at least one of R 2 , R 3 , R 4 , R 5 and R 6 is hydroxyl or amino, and R 2 , R 3 , R 4 , R 5 and R 6 are hydrogen, halogen, lower alkyl, lower alkoxy, amino, substituted amino, hydroxy, carboxyl, or sulfo, or R 5 and R 6 together form a ring, comprising establishing reaction system containing the compound to be assayed and a coupler and an oxidase that consumes O 2 and effects an exzymatic oxidative condensation of said compound to be assayed and said coupler, simultaneously generating a pigment, without the formation of any H 2 O 2 ; and measuring a detectable change in said reaction system to assay said compound.
Claim 2
Independent Claim
143 words
A method for assaying a compound of the formula ##STR10## wherein R 1 is hydroxyl or amino, or hydrogen if at least one of R 2 , R 3 , R 4 , R 5 and R 6 is hydroxyl or amino, and R 2 , R 3 , R 4 , R 5 and R 6 are hydrogen, halogen, lower alkyl, lower alkoxy, amino, substituted amino, hydroxy, carboxyl, or sulfo, or R 5 and R 6 together form a ring, comprising establishing reaction system containing the compound to be assayed and a coupler and an oxidase that consumes O 2 and effects an exzymatic oxidative condensation of said compound to be assayed and said coupler, simultaneously generating a pigment, without the formation of any H 2 O 2 ; and measuring a detectable change in said reaction system to assay said compound.
Claim 3
Dependent Claim
114 words
An assay method according to claim 1, in which R 1 is amino, or hydrogen if at least one of R 2 , R 3 , R 4 , R 5 and R 6 is amino, and R 2 , R 3 , R 4 , R 5 and R 6 have the same meanings as before, said coupler being a compound of the formula ##STR11## wherein R 7 is hydroxyl, amino or substituted amino, and R 8 , R 9 , R 10 , R 11 and R 12 are hydrogen, halogen, lower alkyl, lower alkoxy, amino, substituted amino, hydroxyl, carboxyl or sulfo, or R 11 and R 12 together form a ring.
Claim 4
Dependent Claim
113 words
An assay method according to claim 1, in which R 1 is hydroxyl, or hydrogen if at least one of R 2 , R 3 , R 4 , R 5 and R 6 is hydroxyl, and R 2 , R 3 , R 4 , R 5 and R 6 have the same meanings as before, said coupler being a compound of the formula ##STR12## wherein R 13 is amino or substituted amino, and R 14 , R 15 , R 16 , R 17 and R 18 are hydrogen, halogen, lower alkyl, lower alkoxy, amino substituted amino, hydroxyl, carboxyl or sulfo, or R 17 and R 18 together form a ring.
Claim 5
Dependent Claim
24 words
An assay method according to claim 1, wherein said oxidase is an oxidase for an aromatic ring compound having a hydroxyl or amino substituent.
Claim 6
Dependent Claim
16 words
An assay method according to claim 4, wherein said oxidase is ascorbate oxidase, laccase or tyrosinase.
Claim 7
Dependent Claim
20 words
An assay method according to claim 1, wherein said detectable change is the amount of consumed oxygen or generated pigment.
Claim 8
Dependent Claim
12 words
An assay method according to claim 6, and performing said measurement electrochemically.
Claim 9
Dependent Claim
15 words
An assay method according to claim 7, and performing said measurement by an oxygen electrode.
Claim 10
Dependent Claim
27 words
An assay method according to claim 6, and performing said measurement of the amount of generated pigment by a colorimetric assay at a specific absorption wavelength thereof.
Claim 11
Dependent Claim
25 words
An assay method according to claim 1, wherein said compound to be assayed is generated from a synthetic substrate by the action of a hydrolase.
Claim 12
Dependent Claim
74 words
An assay method according to claim 10, wherein said hydrolase is a peptidase or protease and the synthetic substrate is a compound of the formula ##STR13## wherein one of R' 1 and R' 2 is R 19 CO--NH-- and the other is hydrogen, in which R 19 CO-- is an amino acid residue or peptide residue, and R 3 , R 4 , R 5 and R 6 have the same meanings as before.
Claim 13
Dependent Claim
72 words
An assay method according to claim 10, wherein said hydrolase is glycosidase and the synthetic substrate is a compound of the formula ##STR14## wherein one of R" 1 and R" 2 is (S) n --O-- and the other is hydrogen, in which S is a monosaccharide unit and n is integer greater than 1, and R 3 , R 4 , R 5 and R 6 have the same meanings as before.
Claim 14
Dependent Claim
69 words
An assay method according to claim 10, wherein said hydrolase is a phosphatase and the synthetic substrate is a compound of the formula ##STR15## wherein one of R"' 1 and R"' 2 is ##STR16## and the other is hydrogen, in which R 20 ##STR17## or a substituted or unsubstituted glycerol residue, and R 3 , R 4 , R 5 and R 6 have the same meanings as before.
Claim 15
Dependent Claim
71 words
An assay method according to claim 10, wherein said hydrolase is an esterase or lipase, and the synthetic substrate is a compound of the formula ##STR18## wherein one of R"" 1 and R"" 2 is R 21 CO--O-- and the other is hydrogen, in which R 21 CO-- is a fatty acid residue, and R 3 , R 4 , R 5 and R 6 have the same meanings as before.
Claim 16
Independent Claim
174 words
A method for assaying a hydrolase, comprising the steps of: generating from a synthetic substrate by the action of said hydrolase a compound of the formula ##STR19## wherein R 1 is hydroxyl or amino, or hydrogen if at least one of R 2 , R 3 , R 4 , R 5 and R 6 is hydroxyl or amino, and R 2 , R 3 , R 4 , R 5 and R 6 are hydrogen, halogen, lower alkyl, lower alkoxy, amino, substituted amino, hydroxy, carobxyl, or sulfo, or R 5 and R 6 together form a ring; establishing a reaction system containing a said compound and a coupler and an oxidase that consumes O 2 and effects an enzymatic oxidative condensation of said compound and said coupler, simultaneously generating a pigment, without the formation of any H 2 O 2 ; measuring a detectable change in said reaction system to assay said compound; and relating the assay of said compound to said hydrolase consumed in generating said compound, thereby to assay said hydrolase.
Claim 17
Independent Claim
174 words
A method for assaying a hydrolase, comprising the steps of: generating from a synthetic substrate by the action of said hydrolase a compound of the formula ##STR19## wherein R 1 is hydroxyl or amino, or hydrogen if at least one of R 2 , R 3 , R 4 , R 5 and R 6 is hydroxyl or amino, and R 2 , R 3 , R 4 , R 5 and R 6 are hydrogen, halogen, lower alkyl, lower alkoxy, amino, substituted amino, hydroxy, carobxyl, or sulfo, or R 5 and R 6 together form a ring; establishing a reaction system containing a said compound and a coupler and an oxidase that consumes O 2 and effects an enzymatic oxidative condensation of said compound and said coupler, simultaneously generating a pigment, without the formation of any H 2 O 2 ; measuring a detectable change in said reaction system to assay said compound; and relating the assay of said compound to said hydrolase consumed in generating said compound, thereby to assay said hydrolase.
Claim 18
Dependent Claim
19 words
An assay method according to claim 15, wherein the said hydrolase assay is an assay of piptidase or protease.
Claim 19
Dependent Claim
17 words
An assay method according to claim 15, wherein the said hydrolase assay is an assay of glycosidase.
Claim 20
Dependent Claim
17 words
An assay method according to claim 15, wherein the said hydrolase assay is an assay of phosphatase.
Claim 21
Dependent Claim
19 words
An assay method according to claim 15, wherein the said hydrolase assay is an assay of esterase or lipase.
References Cited (Backward Citations)
US4177109A
Cited
US4209459A
Cited
US4384041A
Cited
GB2103607A
Cited
US4472499A
Cited
US4472499B1
Cited
US4529709A
Cited
DE3331588A1
Cited
EP0108526A2
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1990-07-20 | 1991-01-19 | 1991-07-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-07-20 | 1995-01-19 | 1995-07-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-07-20 | 1999-01-19 | 1999-07-21 | $7700.00 | Window Closed / Lapsed |