Description:
Reference #: 1752
The University of South Carolina is offering licensing opportunities for Arginase Assay Utilizing a Fluorometric-Based Coupled Enzyme System.
Background:
The enzyme arginase is typically assayed for activity by either colorimetry, fluorimetry, or detecting for the concentration of radiolabeled isotopes. In most of the methodologies that are available, high-throughput screening for the evaluation of potent arginase inhibitors is limited. The traditional assays have poor suitability for rapid, automated high-throughput screening of large compound libraries. Colorimetric/spectrophotometric methods require a lengthy color development step and the use of high acidity (e.g., using ninhydrin or diacetyl monoxime to detect urea). Radioisotopes have the drawback of working with an extra level of safety precautions and an increase in the time the experiment can be completed. A modern fluorometric method uses proprietary bioreagents in their assay kit, thus disguising the chemical reactions taking place. These drawbacks lead to poor suitability for high-throughput screening because they lead to low reproducibility, high reagent cost, long turnaround times, or hazardous conditions.
Invention Description:
This invention is a rapid, novel, and sensitive fluorometric-based kinetics assay procedure that detects arginase activity. A urease and glutamate coupled enzyme system is used. Human arginase I (HAI) was selected as a model system to represent the arginase reaction, in which L-arginine is converted into products L-ornithine and urea. The enzymes chosen use NADPH as a substrate along with ammonia and one other substrate to form NADP+, a fluorometrically silent compound. Loss of fluorescence with respect to NADPH is characterized by arginase activity.
Potential Applications:
Pharmaceutical industry, medical industry, biotechnology industry
Advantages and Benefits:
This method can cut down on assay times and minimizes cross-reactivity between the assay and the tested compounds.