Optics: measuring and testing – By dispersed light spectroscopy – With sample excitation
Reexamination Certificate
1997-09-18
2001-01-09
Evans, F. L. (Department: 2877)
Optics: measuring and testing
By dispersed light spectroscopy
With sample excitation
C356S417000, C356S436000, C356S442000, C250S458100, C250S459100
Reexamination Certificate
active
06172750
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to the field of scientific research instrumentation and, in particular, to fluorescence spectroscopy. The present invention includes a method that allows the utilization of fluorescent tracers in techniques where previously only radioactive tracers could be used. For example, the present invention is applied in fluorescent techniques to test for the presence of an enzyme inhibitor in a crude extract or otherwise spectrally interfering substance.
2. Description of the Prior Art
There is no known method for spectral interference correction applied to fluorescence spectroscopy, except for applicant's copending application titled: A Florescent Assay and Method That Corrects For Spectral Interference, which is incorporated by reference, that relates to a method that incorporates a separate or additional fluorescent label into the fluorescent assay. The additional label is unique in that it acts as a measure of interference caused by a colored or otherwise spectrally interfering compound. The present invention, however, eliminates the use of the additional label and, is applied to the primary fluorescent label in the assay.
An article entitled: Use of Fluorescent Cholesteryl Ester Microemulsions In Cholesteryl Ester Transfer Protein Assays by Charles L. Bisgaier, Laura Minton, Arnold D. Essenberg, Andrew White, and Reynold Homen published in the Journal of Lipid Research, Volume 34, 1993, discloses a fluorescent method that measures the activity of lipid transfer protein. The authors point out the limited use of their method due to spectral interference caused by the presence of colored compounds.
The present method is readily applicable to any fluorimeter and restores accuracy to fluorometric measurements in the presence of compounds that may otherwise prevent accurate measurement.
The present invention accomplishes this without the use additional substrates, utilizing standard laboratory equipment.
SUMMARY OF THE INVENTION
It is a principal object of this invention to provide a method of data analysis for accurate determination of enzyme activity utilizing fluorescence, in a sample that would otherwise yield inaccurate results due to spectrally interfering components.
It is another object of the present invention to provide such a method for individuals in the field for the purpose of determining the activity of an enzyme in the presence of a potential inhibitor.
It is still another object of the present invention to provide such a device that is accurate and without utilization of radioisotopes.
It is yet another object of the present invention to provide a new method to measure conventional spectrophotometric assays in a fluorimeter instead of an absorption spectrometer and thus increasing sensitivity of the conventional assay.
To the accomplishments of the foregoing objects and advantages, the present invention, in brief summary, comprises a method that accurately determines the change in fluorescence of a fluorescent measurement without regard to the presence of colored or otherwise interfering factors.
REFERENCES:
patent: 5104228 (1992-04-01), Baillie
patent: 5173434 (1992-12-01), Morris et al.
patent: 5303026 (1994-04-01), Strobl et al.
patent: 5400137 (1995-03-01), Winslow et al.
patent: 5459567 (1995-10-01), Brocia
patent: 5565328 (1996-10-01), Bascomb et al.
Agris Cheryl H.
Evans F. L.
O'Gorman Carmella
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