Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1995-12-01
1998-09-08
Wilson, James O.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 4, 435 772, 435 15, 435 24, 548126, 548405, G01N 33573, G01N 3353, C12Q 100
Patent
active
058043950
ABSTRACT:
Fluorescent-labeled substrates are provided for fluorescence polarization says of enzymes. These substrates are proteins labeled with derivatives of BODIPY.RTM., 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene. The BODIPY.RTM. fluorescent tag of the present invention is pH independent, and can be used over a pH range of from about 2 to about 11. Thus one can assay, in real time, enzymes with pH maxima at pH below 7 using fluorescence polarization methodology, which could not be done with fluorescein derivatives. Different enzymes can be compared using the same BODIPY.RTM. conjugate by merely changing the buffer system which changes the pH conditions.
Fluorescence polarization assays of enzyme activity can be performed in the presence of whole bacteria and other finely suspended particles, such as might be present in tissue homogenates or cellular material. This is particularly useful for chairside assays on dental plaque or clinical assays on bacteria or tissue or exudates.
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Fa et al., Biochemistry, vol. 34, No. 42, 13833-13840, Oct. 1995.
Jolley Michael Ernest
Schade Sylvia Zottu
Spevack A. D.
The United States of America as represented by the Secretary of
Wilson James O.
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