Amino acid substituted-cresyl violet, synthetic fluorogenic substrates for the analysis of agents in individual in vivo cells or tissue

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United States of America Patent

PATENT NO 6235493
SERIAL NO

09130193

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Abstract

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The present invention concerns a method to detect the presence of an enzyme in in vivo or in vitro tissue or cell, which method comprises: (a) obtaining a tissue or cell sample to be analyzed; (b) contacting the tissue or cell sample with a substrate of the structure selected from the group consisting of: ##STR1## X=H or one or more natural or synthetic amino acids with or without amino blocking groups, Y=H or one or more natural or synthetic amino acids with or without amino blocking groups, wherein X and Y are the same or different and are amino acid sequences of between about 1 to 1,000,000 amino acids wherein each amino acid is the same or a different amino acid, with the proviso that at least one of X or Y is at least one amino acid; (c) when an enzyme is present in the tissue or cell sample which degrades X, Y and combinations thereof, fluorescent cresyl violet is released in the tissue sample producing a color change; (d) detecting the presence and amount of the enzyme present by the detection and quantification of the fluorescence produced; and (e) optionally comparing the fluorescence to a pre-calibrated fluorescence scale to quantify the fluorescence present. A diagnostic kit for use and a method to prepare amino acid cresyl violet derivatives are described.

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Patent Owner(s)

  • MP BIOMEDICALS, INC.;REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE

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Inventor(s)

Inventor Name Address # of filed Patents Total Citations
Bissell, Eugene R Alamo, CA 6 150
Smith, Robert F Livermore, CA 119 1725

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