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

US6235493A · kind A · utility

Assignee

• The Regents of the University of California · US

Inventors

• Eugene R. Bissell · Alamo, US
• Robert F. Smith · Newbury Park, US

Key dates

• Filing date: Aug 5, 1998
• Grant date: May 22, 2001
• Expiry date: Aug 5, 2018

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2337/00
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

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: PA1 (a) obtaining a tissue or cell sample to be analyzed; PA1 (b) contacting the tissue or cell sample with a substrate of the structure selected from the group consisting of: ##STR1## PA1 X=H or one or more natural or synthetic amino acids with or without amino blocking groups, PA1 Y=H or one or more natural or synthetic amino acids with or without amino blocking groups, PA1 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; PA1 (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; PA1 (d) detecting the presence and amount of the enzyme present by the detection and quantification of the fluorescence produced; and PA1 (e) optionally comparing the fluorescence to a pre-calibrated fluorescence scale to quantify the fl…