US 11,874,281 B2
Assays for detecting modified compounds
Elazar Rabbani, New York, NY (US); Jannis G. Stavrianopoulos, Bay Shore, NY (US); Joshua Rabbani, New York, NY (US); and Praveen Pande, Holbrook, NY (US)
Assigned to Enzo Life Sciences, Inc., Farmingdale, NY (US)
Filed by Enzo Life Sciences, Inc., Farmingdale, NY (US)
Filed on Oct. 14, 2021, as Appl. No. 17/501,143.
Application 14/296,756 is a division of application No. 12/806,950, filed on Aug. 24, 2010, granted, now 8,778,614, issued on Jul. 15, 2014.
Application 17/501,143 is a continuation of application No. 16/225,669, filed on Dec. 19, 2018, granted, now 11,181,528.
Application 16/225,669 is a continuation of application No. 15/195,343, filed on Jun. 28, 2016, abandoned.
Application 15/195,343 is a continuation of application No. 14/296,756, filed on Jun. 5, 2014, granted, now 9,404,143, issued on Aug. 2, 2016.
Prior Publication US 2022/0034897 A1, Feb. 3, 2022
Int. Cl. G01N 33/68 (2006.01); G01N 33/92 (2006.01); C12Q 1/48 (2006.01)
CPC G01N 33/6803 (2013.01) [C12Q 1/485 (2013.01); G01N 33/92 (2013.01); C12Y 207/01091 (2013.01); C12Y 207/11001 (2013.01); G01N 2405/08 (2013.01); G01N 2410/00 (2013.01); G01N 2440/14 (2013.01); G01N 2440/30 (2013.01)] 7 Claims
 
1. 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.