Methods for the detection and determination of ligands
First Claim
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1. 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.
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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.
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Citations
28 Claims
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1. A method of determining a ligand in a liquid medium suspected of containing same, which method comprises:
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(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. - View Dependent Claims (2, 3, 4, 5, 6, 25)
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7. A method for determining a ligand in a liquid medium suspected of containing same, which method comprises:
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(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. - View Dependent Claims (8, 9, 10, 11, 12, 13, 14, 15, 26)
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16. A method for determining a ligand in a liquid medium suspected of containing same, which method comprises:
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(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. - View Dependent Claims (17, 18, 19, 20, 21, 27)
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22. A method for determining a ligand in a liquid medium suspected of containing same, which method comprises:
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(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. - View Dependent Claims (23, 24, 28)
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Specification