US4228237A
Expired / Lapsed
US
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Methods for the detection and determination of ligands
Assignee: CNA Holdings LLC
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Filing: 1978-09-21
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Granted: 1980-10-14
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Expires: 1998-09-21
Abstract
Abstract A method for determining the presence of a ligand in a liquid medium which utilizes enzyme labeled avidin and biotin labeled reagent in a specific binding process wherein the ligand to be detected is contacted with an insoluble phase containing specific binding substance for the ligand. Various protocols may be used to assay the resulting enzyme activity which is related to the amount of ligand in the liquid medium.
Inventors
- 👤 Richard C. Hevey
- 👤 Mark K. Malmros
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 14:03
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54306; G01N33/531; G01N33/532; Y; Y10; Y10S; Y10S436/00; Y10S436/807; Y10S436/81; Y10S436/815
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
100 words
A method of determining a ligand in a liquid medium suspected of containing same, which method comprises: (a) providing an insoluble phase containing a specific binding substance for said ligand; (b) incubating said insoluble phase with the following reagents: (i) liquid medium suspected of containing said ligand; (ii) biotin labeled specific binding substance for said ligand; and (iii) enzyme labeled avidin; (c) separating unreacted reagents from said insoluble phase after incubation; and (d) determining the enzyme activity of either said insoluble phase or separated unreacted reagent whereby said activity is related to the amount of ligand in said liquid medium.
Claim 2
Independent Claim
100 words
A method of determining a ligand in a liquid medium suspected of containing same, which method comprises: (a) providing an insoluble phase containing a specific binding substance for said ligand; (b) incubating said insoluble phase with the following reagents: (i) liquid medium suspected of containing said ligand; (ii) biotin labeled specific binding substance for said ligand; and (iii) enzyme labeled avidin; (c) separating unreacted reagents from said insoluble phase after incubation; and (d) determining the enzyme activity of either said insoluble phase or separated unreacted reagent whereby said activity is related to the amount of ligand in said liquid medium.
Claim 3
Dependent Claim
26 words
A method according to claim 1 wherein step (b) comprises incubating said insoluble phase with reagents (i), (ii) and (iii) in the presence of one another.
Claim 4
Dependent Claim
56 words
A method according to claim 1 wherein step (b) comprises incubating said insoluble phase with reagents (i), (ii), and (iii) in the order indicated; step (c) comprises separating unreacted reagent from said insoluble phase after each incubation; and step (d) comprises determining the enzyme activity of either said insoluble phase or separated unreacted enzyme labeled avidin.
Claim 5
Dependent Claim
12 words
A method according to claim 1 wherein said ligand is an antigen.
Claim 6
Dependent Claim
13 words
A method according to claim 4 wherein said antigen is alpha 1 fetoprotein.
Claim 7
Dependent Claim
12 words
A method according to claim 1 wherein said ligand is an antibody.
Claim 8
Independent Claim
101 words
A method for determining a ligand in a liquid medium suspected of containing same, which method comprises: (a) providing an insoluble phase containing a specific binding substrate for said ligand; (b) incubating said insoluble phase with the following reagents: (i) liquid medium suspected of containing said ligand; and (ii) biotin labeled specific binding substance for said ligand bound to enzyme labeled avidin; (c) separating unreacted reagents from said insoluble phase after incubation; and (d) determining the enzyme activity of either said insoluble phase or separated unreacted reagent whereby said activity is related to the amount of ligand in said liquid medium.
Claim 9
Independent Claim
101 words
A method for determining a ligand in a liquid medium suspected of containing same, which method comprises: (a) providing an insoluble phase containing a specific binding substrate for said ligand; (b) incubating said insoluble phase with the following reagents: (i) liquid medium suspected of containing said ligand; and (ii) biotin labeled specific binding substance for said ligand bound to enzyme labeled avidin; (c) separating unreacted reagents from said insoluble phase after incubation; and (d) determining the enzyme activity of either said insoluble phase or separated unreacted reagent whereby said activity is related to the amount of ligand in said liquid medium.
Claim 10
Dependent Claim
25 words
A method according to claim 7 wherein step (b) comprises incubating said insoluble phase with reagents (i) and (ii) in the presence of one another.
Claim 11
Dependent Claim
62 words
A method according to claim 7 wherein step (b) comprises incubating said insoluble phase with reagents (i) and (ii) in the order indicated; step (c) comprises separating unreacted reagent from said insoluble phase after each incubation; and step (d) comprises determining the enzyme activity of either said insoluble phase or separated unreacted biotin labeled specific binding substance bound to enzyme labeled avidin.
Claim 12
Dependent Claim
12 words
A method according to claim 7 wherein said ligand is an antigen.
Claim 13
Dependent Claim
12 words
A method according to claim 10 wherein said antigen is human IgE.
Claim 14
Dependent Claim
13 words
A method according to claim 10 wherein said antigen is alpha 1 fetoprotein.
Claim 15
Dependent Claim
12 words
A method according to claim 7 wherein said ligand is an antibody.
Claim 16
Dependent Claim
13 words
A method according to claim 13 wherein said antibody is humann anti-rubella IgG.
Claim 17
Dependent Claim
32 words
A method according to claim 14 wherein the specific binding substance contained in said insoluble phase is rubella antigen and the specific binding substance contained in reagent (ii) is anti human IgG.
Claim 18
Independent Claim
101 words
A method for determining a ligand in a liquid medium suspected of containing same, which method comprises: (a) providing an insoluble phase containing a specific binding substance for said ligand; (b) incubating said insoluble phase with the following reagents: (i) (A) liquid medium suspected of containing said ligand and (B) a known quantity of biotin labeled ligand; and (ii) enzyme labeled avidin; (c) separating unreacted reagents from said insoluble phase after incubation; and (d) determining the enzyme activity of either said insoluble phase or separated unreacted reagent whereby said activity is related to the amount of ligand in said liquid medium.
Claim 19
Independent Claim
101 words
A method for determining a ligand in a liquid medium suspected of containing same, which method comprises: (a) providing an insoluble phase containing a specific binding substance for said ligand; (b) incubating said insoluble phase with the following reagents: (i) (A) liquid medium suspected of containing said ligand and (B) a known quantity of biotin labeled ligand; and (ii) enzyme labeled avidin; (c) separating unreacted reagents from said insoluble phase after incubation; and (d) determining the enzyme activity of either said insoluble phase or separated unreacted reagent whereby said activity is related to the amount of ligand in said liquid medium.
Claim 20
Dependent Claim
25 words
A method according to claim 16 wherein step (b) comprises incubating said insoluble phase with reagents (i) and (ii) in the presence of one another.
Claim 21
Dependent Claim
55 words
A method according to claim 16 wherein step (b) comprises incubating said insoluble phase with reagents (i) and (ii) in the order indicated; step (c) comprises separating unreacted reagent from said insoluble phase after each incubation; and step (d) comprises determining the enzyme activity of either said insoluble phase or separated unreacted enzyme labeled avidin.
Claim 22
Dependent Claim
12 words
A method according to claim 16 wherein said ligand is an antigen.
Claim 23
Dependent Claim
12 words
A method according to claim 19 wherein said antigen is human IgG.
Claim 24
Dependent Claim
12 words
A method according to claim 16 wherein said ligand is an antibody.
Claim 25
Independent Claim
100 words
A method for determining a ligand in a liquid medium suspected of containing same, which method comprises: (a) providing an insoluble phase containing a specific binding substance for said ligand; (b) incubating said insoluble phase with a reagent comprising (A) liquid medium suspected of containing said ligand and (B) a known quantity of biotin labeled ligand bound to enzyme labeled avidin; (c) separating unreacted reagent from said insoluble phase after incubation; and (d) determining the enzyme activity of either said insoluble phase or separated unreacted reagent whereby said activity is related to the amount of ligand in said liquid medium.
Claim 26
Independent Claim
100 words
A method for determining a ligand in a liquid medium suspected of containing same, which method comprises: (a) providing an insoluble phase containing a specific binding substance for said ligand; (b) incubating said insoluble phase with a reagent comprising (A) liquid medium suspected of containing said ligand and (B) a known quantity of biotin labeled ligand bound to enzyme labeled avidin; (c) separating unreacted reagent from said insoluble phase after incubation; and (d) determining the enzyme activity of either said insoluble phase or separated unreacted reagent whereby said activity is related to the amount of ligand in said liquid medium.
Claim 27
Dependent Claim
12 words
A method according to claim 22 wherein said ligand is an antigen.
Claim 28
Dependent Claim
12 words
A method according to claim 22 wherein said ligand is an antibody.
Claim 29
Independent Claim
44 words
A method according to claims 1, 2, 3, 4, 5 or 6 wherein said insoluble phase, said biotin labeled specific binding substance and said enzyme labeled avidin are each present during incubation in excess of the total amount of ligand in the liquid medium.
Claim 30
Independent Claim
48 words
A method according to claims 7, 8, 9, 10, 11, 12, 13, 14, or 15 wherein said insoluble phase and said biotin labeled specific binding substance bound to enzyme labeled avidin are each present during incubation in excess of the total amount of ligand in the liquid medium.
Claim 31
Independent Claim
58 words
A method according to claims 16, 17, 18, 19, 20 or 21 wherein said ligand to be determined, said biotin labeled ligand and said enzyme labeled avidin are each present during incubation in excess of said insoluble phase, the quantity of biotin labeled ligand not exceeding the smallest quantity of ligand to be determined in the liquid medium.
Claim 32
Independent Claim
61 words
A method according to claims 22, 23 or 24 wherein said ligand to be determined and said biotin labeled ligand bound to enzyme labeled avidin are each present during incubation in excess of said insoluble phase, the quantity of biotin labeled ligand bound to enzyme labeled avidin not exceeding the smallest quantity of ligand to be determined in the liquid medium.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-10-14 | 1984-04-14 | 1984-10-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-10-14 | 1988-04-14 | 1988-10-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-10-14 | 1992-04-14 | 1992-10-14 | $7700.00 | Window Closed / Lapsed |