Patent US3975237A successfully refreshed from Google Patents.
US3975237A
Expired / Lapsed
US
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Compounds for enzyme amplification assay - - ecgonine analogs
Assignee: Syva Co
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Filing: 1974-06-20
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Granted: 1976-08-17
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Expires: 1994-06-20
Abstract
Abstract Novel biological assay method for determining the presence enzyme a specific organic material by employing a modified enayme for amplification. By employing receptors specific for one or a group of materials (hereinafter referred to as "ligands") and binding an enzyme to the ligand or ligand counterfeit to provide an "enzyme-bound-ligand", an extremely sensitive method is provided for assaying for ligands. The receptor when bound to the enzyme-bound-ligand substantially inhibits enzymatic activity, providing for different catalytic efficiencies of enzyme-bound-ligand and enzyme-bound-ligand combined with receptor. The receptor, ligand and enzyme-bound-ligand are combined in an arbitrary order and the effect of the presence of ligand on enzymatic activity determined. Various protocols may be used for assaying for enzymatic activity and relating the result to the amount of ligand present.
Inventors
- 👤 Kenneth E. Rubenstein
- 👤 Edwin F. Ullman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 16:51
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/535; G01N33/94; G01N33/946; G01N33/948; G01N33/9486; Y; Y10; Y10S; Y10S435/00; Y10S435/964
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
98 words
An enzyme-bound-ligand of the formula: ##EQU9## wherein: any one of the W groups can be --X* or an H of any of the W groups may be replaced by --X*, wherein --X* is a bond or a linking group; A* is an enzyme bonded at other than its reactive site, having n ligands, wherein n is in the range of 1 to the molecular weight of A* divided by 2,000; W 55 is hydroxy, methoxy, amino or methylamino; W 56 is hydrogen or benzoyl; and W 57 is hydrogen or alkyl of from 1 to 3 carbon atoms.
Claim 2
Dependent Claim
98 words
An enzyme-bound-ligand of the formula: ##EQU9## wherein: any one of the W groups can be --X* or an H of any of the W groups may be replaced by --X*, wherein --X* is a bond or a linking group; A* is an enzyme bonded at other than its reactive site, having n ligands, wherein n is in the range of 1 to the molecular weight of A* divided by 2,000; W 55 is hydroxy, methoxy, amino or methylamino; W 56 is hydrogen or benzoyl; and W 57 is hydrogen or alkyl of from 1 to 3 carbon atoms.
Claim 3
Dependent Claim
101 words
An enzyme-bound-ligand of the formula: ##EQU10## wherein: one of W 56 ' and W 57 ' is --X**; when other than --X**, W 56 ' is hydrogen or benzoyl; and W 57 ' is methyl; W 55 ' is hydroxy or methoxy; X** is ##SPC34## wherein: Z a is methylene or carbonyl; or --Z--CO--, or --Z--CNH--, wherein Z is hydrocarbylene of from 1 to 7 carbon atoms; A** is an enzyme having n' ligands, wherein n' is in the range of 1 to 20, and wherein the enzyme is of a molecular weight in the range of about 10,000 to 300,000.
Claim 4
Dependent Claim
101 words
An enzyme-bound-ligand of the formula: ##EQU10## wherein: one of W 56 ' and W 57 ' is --X**; when other than --X**, W 56 ' is hydrogen or benzoyl; and W 57 ' is methyl; W 55 ' is hydroxy or methoxy; X** is ##SPC34## wherein: Z a is methylene or carbonyl; or --Z--CO--, or --Z--CNH--, wherein Z is hydrocarbylene of from 1 to 7 carbon atoms; A** is an enzyme having n' ligands, wherein n' is in the range of 1 to 20, and wherein the enzyme is of a molecular weight in the range of about 10,000 to 300,000.
Claim 5
Dependent Claim
12 words
An enzyme-bound-ligand according to claim 1, wherein said enzyme is an oxidoreductase.
Claim 6
Dependent Claim
11 words
An enzyme-bound-ligand according to claim 1, wherein said enzyme is hydrolase.
Claim 7
Dependent Claim
21 words
An enzyme-bound-ligand according to claim 1 which is para-diazobenzoylecgonine conjugate to lysozyme, having from 2 to 4 of said ecgonine groups.
Claim 8
Dependent Claim
22 words
An enzyme-bound-ligand according to claim 1 which is para-liazobenzoylecgonine conjugate to malate dehydrogenase, having from 2 to 22 of said ecgonine groups.
Claim 9
Dependent Claim
23 words
An enzyme-bound-ligand according to claim 1, which is para-diazobenzoylecgonine conjugate to glucose 6-phosphate dehydrogenase, having from 2 to 22 of said ecgonine groups.
Claim 10
Dependent Claim
16 words
An enzyme-bound-ligand according to claim 1, which is para-(3-ecgoninyloxvcarbonylphenyl)thiocarbamic acid conjugate to an oxidoreductase or hydrolase.
Claim 11
Dependent Claim
19 words
An enzyme-bound-ligand according to claim 1, which is para-(3-eogoninyloxycarbonylphenyl)thiocarbamic acid conjugate to lysozyme, malate dehydrogenase or glucose 6-phosphate dehydrogenase.
Claim 12
Dependent Claim
18 words
An enzyme-bound-ligand according to claim 1, which is para-chloroacetamidebenzoylecgonine conjugate to lysozyme, malate dehydrogenase, or glucose 6-phosphate dehydrogenase.
References Cited (Backward Citations)
US3850752A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1979-08-17 | 1980-02-16 | 1980-08-17 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1983-08-17 | 1984-02-16 | 1984-08-17 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1987-08-17 | 1988-02-16 | 1988-08-17 | $7700.00 | Window Closed / Lapsed |