Double receptor fluorescent immunoassay

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23230B, 1951035R, 260112B, 25230116, G01N 2100, G01N 3316

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active

039989437

ABSTRACT:
A sensitive method for determining qualitatively and quantitatively the presence of a wide variety of physiologically active organic compounds (ligand) is provided. The method employs a reagent which involves bonding a compound having structural similarity to the compound to be determined (ligand analog) to a fluorescing compound. The unknown compound is referred to as a ligand, the conjugate of the structurally similar compound and fluorescer is referred to as ligand analog-fluorescer, and compounds which recognize a specific structure and bind to such structure are referred to as receptors and are normally antibodies.
The fluorescer which is chosen will have either a change in quantum yield or a change in its emisson and/or absorption spectra or all of them, when bound to antibody, as compared to being unbound. For the purposes of the assay, all that is required is that there be a change in the emisson intensity at some wavelength or band of wavelengths.
The rate at which fluorescer antibody binds to the fluorescer portion of the ligand anaog-fluorescer or the amount of fluorescer antibody bound to the fluorescer portion of the ligand analog-fluorescer at equilibrium will be related to the amount of ligand antibody bound to the ligand analog portion of the ligand analog-fluorescer. Therefore, by combining antibodies to both ligand and fluorescer, with ligand analog-fluorescer and an unknown, one can determine the amount of ligand present in the unknown by relating the emission intensity at a particular wavelength or band of wavelengths to standards.

REFERENCES:
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patent: 3709868 (1973-01-01), Spector
patent: 3766162 (1973-10-01), Spector
patent: 3775536 (1973-11-01), Spector
patent: 3789116 (1974-01-01), Kay
patent: 3809782 (1974-05-01), Spector
patent: 3817706 (1974-06-01), Smith
patent: 3901654 (1975-08-01), Gross
patent: 3935074 (1976-01-01), Rubenstein
Chemical Abstracts, 73:64436m (1970).
Chemical Abstracts, 73:48541m (1970).
Chemical Abstracts, 79:101164t (1973).

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