Real time methylumbelliferone-based assay

Details Real time methylumbelliferone-based assay Real time methylumbelliferone-based assay

2005-05-13

2009-02-10

Weber, Jon P (Department: 1657)

Drug, bio-affecting and body treating compositions

Designated organic active ingredient containing

Carbohydrate doai

C438S206000

Reexamination Certificate

active

07488721

ABSTRACT:
A method is provided for determining the activity of an enzyme which releases methylumbelliferone (MU) from an MU-containing substrate wherein the enzyme has a pH optimum below the pKa of MU comprising: contacting a sample suspected of containing the enzyme with the MU-containing substrate at a pH suitable for activity of the enzyme to allow release of MU by the enzyme; contacting the sample with light of a wavelength in the range of about 310 nm to about 350 nm; determining fluorescence produced by the released MU, thereby determining the activity of the enzyme. This real time method provides improved diagnostic methods, for example for diseases associated with an abnormal level of activity of a glycosidase. The real time assay also can be used to screen compounds for their ability to modulate enzyme activity using MU-containing substrates.

REFERENCES:
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patent: 2002/0035072 (2002-03-01), Fan et al.
patent: WO 2004/103368 (2004-12-01), None
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Application Note SAFIRE (2001) TECAN, “Shifts in Emission and Excitation Spectra due to pH Changes”, pp. 1 to 7.

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