Process for conducting fluorescence immunoassays without added l

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or...

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435 5, 435 7, 435 34, 436517, 436540, 436546, 436805, 436815, 436817, 436816, G01N 3354, G01N 3106, G01N 2164

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043680470

ABSTRACT:
A process is presented for conducting fluorescence immunoassays without the use of added labels by utilizing ultraviolet radiation and internal reflection optics to activate fluorescent groups present in the molecules of interest. The assay is accomplished by directing a beam of light having wavelengths in the ultraviolet region to a solid liquid interface which (1) has a material X immobilized thereon, and (2) is contacted with an assay solution containing in unknown Y which contains intrinsic tyrosine, tryptophan, nucleic acid or related fluorescence groups which are activated by wavelengths in the ultraviolet region; X and Y may be in the relationship of antibody and antigen in that one had the ability of recognizing the particular spatial and polar configuration of the other and is attracted to and bound to such configuration, said beam of ultraviolet light being projected under such conditions that there is internal reflection at the interface, and then measuring the amount of fluorescence emitted from the surface of the interface, the amount of the fluorescence emission being a function of the amount of the unknown Y being detected. In the event that Y is not fluorescent, the X-Y sandwich can be further exposed to a solution containing X, to form XYX sandwich. The observed fluorescence of X is proportional to the amount of bound Y which is in turn related to the solution concentration of Y.

REFERENCES:
patent: 3939350 (1976-02-01), Kronick
patent: 4050895 (1977-09-01), Hardy
E. S. West et al., "Textbook of Biochemistry", Fourth Edition, p. 342, Macmillan, New York, 1967.

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