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US 11,874,281 B2Granted
Assays for detecting modified compounds
AssigneeEnzo Life Sciences, Inc.
InventorsElazar Rabbani, Jannis G. Stavrianopoulos, Joshua Rabbani, Praveen Pande
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Abstract
Provided are methods and compositions which are useful for separating, isolating, detecting, and quantifying compounds of interest which have been modified chemically, enzymatically or catalytically from other compounds which have not been so modified. The modifications may take the form of functional groups which are gained, lost or retained by the compounds of interest.
Claims7
A method for detecting or quantifying an enzyme capable of adding a functional group to a compound in a sample, the method comprising the steps of:
- (a) providing:
- (i) a compound which can gain a functional group from an enzyme, the compound being directly covalently bound to at least one non-radioactive signaling moiety and the compound comprising sphingosine;
- (ii) a sample suspected of containing an enzyme that could add a functional group to the compound;
- and
- (iii) a chemical, enzymatic or catalytic source of the functional group which can be gained by the compound;
- and
- (iv) means to separate the compound which has gained the functional group from the compound which has not gained the functional group;
- (b) forming a mixture comprising the compound, the sample and the source of the functional group, and incubating the mixture under conditions suitable for the compound to gain the functional group;
- (c) separating and thereby isolating any compound which has gained the functional group from any compound which has not gained the functional group by precipitating said compound which has gained the functional group from the mixture by the addition of a charged metal ion and a solvent selected from the group consisting of ethanol, methanol, isopropyl alcohol, dioxane, and combinations thereof;
- (d) resolubilizing said compound which has gained the functional group;
- wherein the functional group comprises a phosphate.
8 more elements
Dependencies are inferred from the claim text.