Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase
Patent
1996-02-05
1998-08-18
Leary, Louise N.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving hydrolase
435 23, 435 25, 435 18, 435 4, 435 681, 435968, 536 243, 252582, 252583, C12Q 137, C12Q 126, C12Q 100, G01N 3353
Patent
active
057957290
ABSTRACT:
An energy transfer fluorescent probe for detecting a reagent is provided which includes a fluorescent reporter molecule and a quencher molecule positioned on the probe relative to the reporter molecule such that the quencher molecule quenches the fluorescence of the reporter molecule when in a first state, the quencher molecule being converted by the reagent to a second state which has a reduced ability to quench the reporter molecule. Examples of conversions of the quencher molecule from a first state to a second state include reductions, oxidations, hydrolyses, phosphate cleavages, and the conversion of amides to amines. In one embodiment, the quencher molecule is a substrate for an enzyme which converts the quencher from a first state to a second state. For example, the enzyme may be an reductase, an oxidase, hydrolytic, a peptidase or a phosphorylase. The probe is used to fluorescently detect a reagent in a sample. According to the method, the probe is contacted with the sample containing the reagent wherein the quencher molecule is converted by the reagent from a first state which is able to quench the fluorescence of the reporter molecule to a second state which has a reduced ability to quench the reporter molecule. The reagent is detected by monitoring the fluorescence intensity of the reporter molecule as the quencher molecule is converted from the first state to the second state by the reagent.
REFERENCES:
patent: 4318981 (1982-03-01), Burd et al.
patent: 4699876 (1987-10-01), Libeskind
patent: 4910300 (1990-03-01), Urdea et al.
patent: 5470705 (1995-11-01), Grossman et al.
Leventhal, et al., Insulin-like Growth Factor-II as a Paracrine Growth Factor in Human Neuroblastoma Cells, Experimental Cell Research 221; pp. 179-186 (1995).
Scudiero, et al., Evaluation of a Soluble Tetrazolium/Formazan Assay for Cell Growth and Drug Sensitivity in Culture Using Human and other Tumor Cell Lines, Sep. 1, 1988; Cancer Research; pp. 4827-4833.
Cory, et al., Use of an Aqueous Soluble Tetrazolium/Formazan Assay for Cell Growth Assays in Culture, Cancer Communications, vol. 3, No. 7, 1991, pp. 207-212.
Paull, et al. The Synthesis of XTT: A New Tetrazolium Reagent that is Bioreducible to a Water-Soluble Formazan, Jun. 1988, pp. 911-914.
Haugland, Detecting Enzymatic Activity in Cells Using Fluorogenic Substrates, Biotechnic & Histochemistry, vol. 70, No. 5, 1995, pp. 243-251.
Barltrop, et al., 5-(3-Carboxymethoxyphenyl)-2-(4,5-Dimethylthiazolyl)-3-(4-Sulfophenyl) Tetrazolium, Inner Salt (MTS) and Related Analogs of 3-(4,5-Dimethylthiazolyl(-2,5-Diphenyltetrazolium Bromide (MTT) Reducing to Purple Water-Soluble Formazans as Cell-Viability Indicators, Bioorganic & Medical Chemistry Letters, vol. 1, No. 11, pp. 611-614, 1991.
Biometric Imaging Inc.
Leary Louise N.
LandOfFree
Reductive, energy-transfer fluorogenic probes does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Reductive, energy-transfer fluorogenic probes, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Reductive, energy-transfer fluorogenic probes will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1113712