Heterogeneous immunoassay for digoxin using ouabain as a separation means
AssigneeE. I. Du Pont de Nemours and Company
InventorsWilliam J. Freytag, Shung-Ho Chang
Life of this patent
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Abstract
A heterogeneous immunoassay for digoxin. An excess of labeled anti-digoxin antibody is added to a test sample and the resulting reaction mixture is contacted with a solid phase having ouabain immobilized thereon. All the free, labeled antibody binds to the oubain. The eluted digoxin anti-digoxin antibody complex is measured for label activity. A competitive mode is also disclosed.
Claims20
A noncompetitive immunoassay for the measurement of digoxin in a test sample, said assay comprising the steps of:
- (a) forming a reaction mixture by contacting a molar excess of labeled monovalent or divalent anti-digoxin antibody with the test sample, whereby a fraction of said antibody forms a complex with the digoxin and a fraction remains free;
- and
- (b) contacting the reaction mixture with a solid phase having ouabain immobilized thereon, the ouabain being present in an amount capable of binding all of the free antibody, whereby the free antibody is bound to the solid phase;
- and
- (c) measuring the amount of complex which elutes from the solid phase by measuring the activity of the label.
A competitive immunoassay for the measurement of digoxin in a test sample, said assay comprising the sequential steps of:
- (a) forming a reaction mixture by contacting a sample suspected of containing digoxin with a molar excess of ouabain immobilized on a solid phase;
- (b) contacting the reaction mixture with a labeled, monovalent or divalent anti-digoxin antibody, said antibody being in immunochemical excess over digoxin, but in immunochemical deficiency relative to ouabain;
- (c) allowing a reaction to take place whereby a fraction of the antibody forms a first complex with the digoxin and a second fraction forms a complex with the immobilized ouabain;
- (d) separating the first fraction from the second fraction;
- and
- (e) measuring the amount of label present in either the first or the second fraction.
A test kit for measuring the amount of digoxin in a liquid sample, comprising:
- (a) a labeled anti-digoxin antibody;
- and
- (b) a solid phase having ouabain bound thereto;
- and
- (c) directions for the use of said kit.
Dependencies are inferred from the claim text.