Double-labeled enzymeimmunoassay methods
First Claim
1. A method of performing an immunoassay of a ligand in a liquid sample wherein two independently measureable enzyme labels are separately conjugated to two or more components or populations of components of the assay system and, after completion of the complexing reaction, substantially all of the first enzyme label and a proportion of the second enzyme label are removed from the assay mixture, the proportion of the second label removed being related to the amount of the said ligand and the assay determined from a measurement of said proportion of the second label being normalised by comparison with a measurement of the total first label removed.
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Abstract
The present invention provides a method of performing an immunoassay of a ligand in a liquid sample wherein two independently measureable enzyme labels are separately conjugated to two or more components or populations of components of the assay system and, after completion of the complexing reaction, substantially all of the first enzyme label and a proportion of the second enzyme label are removed from the assay mixture, the proportion of the second label removed being related to the amount of the said ligand and the assay determined from a measurement of said proportion of the second label being normalised by comparison with a measurement of the total first label removed.
In this way, manual pipetting errors and other assay procedural errors can be corrected for.
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Citations
17 Claims
- 1. A method of performing an immunoassay of a ligand in a liquid sample wherein two independently measureable enzyme labels are separately conjugated to two or more components or populations of components of the assay system and, after completion of the complexing reaction, substantially all of the first enzyme label and a proportion of the second enzyme label are removed from the assay mixture, the proportion of the second label removed being related to the amount of the said ligand and the assay determined from a measurement of said proportion of the second label being normalised by comparison with a measurement of the total first label removed.
- 10. A kit of reagents for carrying out a method of immunoassay which comprises, in at least one container, (i) a ligand analog labelled with a first enzyme label and (ii) a reagent comprising antibodies to the ligand under assay labelled with a second enzyme label, said second enzyme being different from said first enzyme and being measurable independently of said first enzyme label.
- 13. A kit of reagents for carrying out a method of immunoassay comprising in at least one container (i) a first population of antibodies to the ligand under assay labelled with a first enzyme label, and (ii) a second population of antibodies to the ligand under assay labelled with a second enzyme label, said second enzyme being different from said first enzyme and being measurable independently of said first enzyme label, said first and second population of antibodies being directed against two different epitopes.
Specification