Patent US3939319A successfully refreshed from Google Patents.
US4067774A
Expired / Lapsed
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Compounds for enzyme amplification assay
Assignee: Siemens Healthcare Diagnostics GmbH Germany
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Filing: 1976-09-13
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Granted: 1978-01-10
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Expires: 1996-09-13
Abstract
Abstract Novel biological assay method for determining the presence of a specific organic material by employing a modified enzyme 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
2026-10-24 00:43
CPC Classifications
C; C07; C07J; C07J43/00; C07J43/003; C07J19/00; C07J41/00; C07J41/0005; C07J41/0016; 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; Y10S930/00; Y10S930/01; Y10S930/26
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
188 words
An enzyme-bound-ligand of the formula: ##STR61##wherein: any one of the W groups other than W 44 and W 45 can be -X* or an H of any of the W groups other than W 44 or W 45 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 40 and W 41 are hydrogen or alkyl of from 1 to 3 carbon atoms; W 42 is hydrogen, alkyl of from 1 to 3 carbon atoms, or may be taken together with W 40 to form a ring having 6 annular members with a nitrogen as the only heteroatom; W 43 is hydrogen, hydroxyl, carbomethoxy, or may be taken together with W 40 to form a morpholine ring, with the proviso that W 43 is carbomethoxy, when W 40 and W 42 are taken together; and W 44 and W 45 are hydrogen, hydroxyl, or alkoxyl of from 1 to 3 carbon atoms.
Claim 2
Dependent Claim
188 words
An enzyme-bound-ligand of the formula: ##STR61##wherein: any one of the W groups other than W 44 and W 45 can be -X* or an H of any of the W groups other than W 44 or W 45 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 40 and W 41 are hydrogen or alkyl of from 1 to 3 carbon atoms; W 42 is hydrogen, alkyl of from 1 to 3 carbon atoms, or may be taken together with W 40 to form a ring having 6 annular members with a nitrogen as the only heteroatom; W 43 is hydrogen, hydroxyl, carbomethoxy, or may be taken together with W 40 to form a morpholine ring, with the proviso that W 43 is carbomethoxy, when W 40 and W 42 are taken together; and W 44 and W 45 are hydrogen, hydroxyl, or alkoxyl of from 1 to 3 carbon atoms.
Claim 3
Dependent Claim
141 words
An enzyme-bound-ligand of the formula: ##STR62##wherein: any one of the W groups other than W 46' can be -X* or an H of any of the W groups other than W 46' 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 40' and W 41' are hydrogen or alkyl of from 1 to 3 carbon atoms; W 42' is hydrogen, methyl, or may be taken together with W 40' to form a piperidine ring; W 43' is hydrogen, hydroxyl, or carbomethoxy with the proviso that W 43' is carbomethoxy when W 42' is taken together with W 40' ; and W 46' is hydrogen.
Claim 4
Dependent Claim
141 words
An enzyme-bound-ligand of the formula: ##STR62##wherein: any one of the W groups other than W 46' can be -X* or an H of any of the W groups other than W 46' 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 40' and W 41' are hydrogen or alkyl of from 1 to 3 carbon atoms; W 42' is hydrogen, methyl, or may be taken together with W 40' to form a piperidine ring; W 43' is hydrogen, hydroxyl, or carbomethoxy with the proviso that W 43' is carbomethoxy when W 42' is taken together with W 40' ; and W 46' is hydrogen.
Claim 5
Dependent Claim
96 words
An enzyme-bound-ligand of the formula: ##STR63##wherein: one of W 40"' , and W 42"' is -X**; when other than -X**, W 40"' is hydrogen; W 42"' is methyl; and W 44"' is hydrogen; W 41"' is hydrogen or methyl; X** is -Z-C═O wherein Z is hydrocarbylene of from 1 to 7 carbon atoms, with the proviso that when W 44"' is -X**, -X** is ##STR64##and A** is an enzyme having n' ligands, wherein n' is in the range of 1 to 20, and said enzyme has a molecular weight in the range of 10,000 to 300,000.
Claim 6
Dependent Claim
96 words
An enzyme-bound-ligand of the formula: ##STR63##wherein: one of W 40"' , and W 42"' is -X**; when other than -X**, W 40"' is hydrogen; W 42"' is methyl; and W 44"' is hydrogen; W 41"' is hydrogen or methyl; X** is -Z-C═O wherein Z is hydrocarbylene of from 1 to 7 carbon atoms, with the proviso that when W 44"' is -X**, -X** is ##STR64##and A** is an enzyme having n' ligands, wherein n' is in the range of 1 to 20, and said enzyme has a molecular weight in the range of 10,000 to 300,000.
Claim 7
Dependent Claim
12 words
An enzyme-bound-ligand according to claim 3, wherein said enzyme is an oxidoreductase.
Claim 8
Dependent Claim
12 words
An enzyme-bound-ligand according to claim 3, wherein said enzyme is a hydrolase.
Claim 9
Dependent Claim
21 words
An enzyme-bound-ligand according to claim 3, which is N-carboxymethylamphetamine conjugate to lysozyme, having from 2 to 4 of said amphetamine groups.
Claim 10
Dependent Claim
22 words
An enzyme-bound-ligand according to claim 3, which is N-carboxymethylamphetamine conjugate to malate dehydrogenase, having from 2 to 22 of said amphetamine groups.
Claim 11
Dependent Claim
23 words
An enzyme-bound-ligand according to claim 3, which is N-carboxymethylamphetamine conjugate to glucose 6-phosphate dehydrogenase, having from 2 to 22 of said amphetamine groups.
References Cited (Backward Citations)
US3878187A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-01-09 | 1981-07-11 | 1982-01-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-01-09 | 1985-07-11 | 1986-01-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-01-09 | 1989-07-11 | 1990-01-10 | $7700.00 | Window Closed / Lapsed |