US2010041016A1PendingUtilityA1
Methods for detecting organisms and enzymatic reactions using raman spectroscopy and aromatic compounds comprising phosphate
Est. expiryOct 17, 2025(expired)· nominal 20-yr term from priority
G01N 33/56938G01N 33/581C12Q 1/28G01N 21/65G01N 33/58G01N 33/56916G01N 33/56983G01N 2333/55G01N 33/56911C12Q 1/42
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Claims
Abstract
The present disclosure provides systems for the rapid and sensitive detection of organisms and molecules in samples. Reactants that produce Raman-active products are used in combination with Raman light scattering. Such compounds may comprise phosphates permitting the detection of phosphatases. The present disclosure can also be used to measure enzyme-kinetics.
Claims
exact text as granted — not AI-modified1 . A method for detecting the activity of at least one enzyme in a sample comprising:
a) preparing a mixture comprising the sample and at least one aromatic compound comprising at least one phosphate group; b) incubating the mixture to form at least one Raman-active product;
i) optionally adding an oxidizing agent; and
ii) optionally adding a base;
c) detecting the at least one Raman-active product with Raman spectroscopy.
2 . The method of claim 1 , wherein the at least one aromatic compound comprises:
wherein
X is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, PO 4 , and COOH;
Y is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, and COOH; and
Z is chosen from H, OH, Cl, Br, NH 2 , SO 3 H, PO 4 , and COOH.
3 . The method of claim 1 , wherein the at least one aromatic compound comprises:
wherein
X is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, PO 4 , and COOH;
Y is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, and COOH; and
Z is chosen from H, OH, Cl, Br, NH 2 , SO 3 H, PO 4 , and COOH.
4 . The method of claim 1 , wherein the at least one aromatic compound comprises:
wherein
X is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, PO 4 , and COOH; and
Y is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, and COOH.
5 . The method of claim 1 , wherein the at least one aromatic compound comprises:
wherein
X is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, PO 4 , and COOH; and
Y is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, and COOH.
6 . The method of claim 1 , wherein the at least one aromatic compound comprises:
wherein each of X, Y, Z, and W are each independently chosen from H and OH.
7 . The method of claim 1 , wherein the at least one enzyme comprises a phosphatase.
8 . The method of claim 7 , wherein the phosphatase is alkaline phosphatase.
9 . The method of claim 8 , wherein the alkaline phosphatase is conjugated to an antibody.
10 . The method of claim 1 , wherein the at least one aromatic compound comprises 4-amino-1-phenyl-1-phosphate.
11 . The method of claim 1 , wherein the at least one aromatic compound comprises 4-hydroxy-1-naphthyl-1-phosphate.
12 . The method of claim 1 , wherein the at least one aromatic compound comprises 4-amino-1-naphthyl-1-phosphate.
13 . The method of claim 1 , wherein the at least one aromatic compound comprises hydroquinone diphosphate.
14 . The method of claim 1 , wherein the base is sodium hydroxide.
15 . The method of claim 1 , wherein the oxidizing agent is sodium metaperiodate.
16 . The method of claim 1 , wherein the Raman spectroscopy is resonant Raman spectroscopy.
17 . A method for detecting at least one target in a sample comprising:
a) preparing a mixture comprising the at least one target; b) incubating the mixture with at least one ligand specific for the at least one target, wherein the at least one ligand comprises a phosphatase; c) providing to the mixture at least one aromatic compound comprising a phosphate; d) incubating the mixture to form at least one Raman-active product;
i) optionally adding an oxidizing agent; and
ii) optionally adding a base; and
e) detecting the at least one Raman-active product with Raman spectroscopy.
18 . The method of claim 17 , wherein the at least one aromatic compound is the aromatic compound of claim 2 .
19 . The method of claim 17 , wherein the at least one aromatic compound is the aromatic compound of claim 3 .
20 . The method of claim 17 , wherein the at least one aromatic compound is the aromatic compound of claim 4 .
21 . The method of claim 17 , wherein the at least one aromatic compound is the aromatic compound of claim 5 .
22 . The method of claim 17 , wherein the at least one aromatic compound is the aromatic compound of claim 6 .
23 . The method of claim 17 , wherein the Raman spectroscopy is resonance Raman Spectroscopy.
24 . The method of claim 17 , wherein the at least one target is an organism.
25 . The method of claim 24 , wherein the organism is chosen from E. coli, Listeria, Salmonella, Vibrio, Camphelbacter, Staphylococcus , HIV, Hepatitis, Adenovirus, Rhino virus, and Human papilloma virus.
26 . The method of claim 17 , wherein the target is chosen from protein, amino acids, nucleic acids, nucleotides, carbohydrates, metabolites, hormones, and metabolic intermediates.
27 . The method of claim 26 , wherein the protein is chosen from IL-2, C-reactive protein, Tumor Necrosis Factor Receptor II, and Human Cardiac Troponin I.
28 . The method of claim 17 , wherein the at least one ligand is an antibody.
29 . A kit for detecting at least one enzyme activity comprising:
a) at least one aromatic compound comprising a phosphate; b) optionally an oxidizing agent; c) optionally a base; and d) optionally suitable buffers for the at least one enzyme.
30 . An aromatic compound comprising:
wherein
X is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, PO 4 , and COOH;
Y is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, COOH; and
Z is chosen from H, OH, Cl, Br, NH 2 , SO 3 H, PO 4 , and COOH.
31 . (canceled)
32 . An aromatic compound comprising:
wherein
X is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, PO 4 , and COOH; and
Y is chosen from H, OH, Cl, Br, NO 2 , NH 2 , SO 3 H, and COOH.
33 - 34 . (canceled)
35 . An aromatic compound comprising:
wherein each of X, Y, Z, and W are each independently chosen from H and OH.Join the waitlist — get patent alerts
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