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US 9,540,702 B2Granted
Stabilization of enzymes with stable coenzymes
AssigneeRoche Diabetes Care, Inc.
InventorsWolfgang Roedel, Carina Horn, Nelli Steinke, Nadine Bucci, Thomas Meier, Rainer Schmuck, Rolf Nagel, Dieter Heindl
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Abstract
Methods for stabilizing an enzyme by storing the enzyme in the presence of a stabilized coenzyme are disclosed. In addition, an enzyme stabilized with a stabilized coenzyme as well as the use thereof in test elements for detecting analytes are also disclosed. Other aspects include unique compositions, methods, techniques, systems and devices involving enzyme stabilization.
Claims11
A method for stabilizing an enzyme, which comprises:
- storing the enzyme in a presence of a stabilized coenzyme, or a salt form thereof, or a reduced form thereof for a period of storage of at least 2 weeks at a temperature of at least 20° C., wherein the enzyme is selected from the group consisting of an alcohol dehydrogenase (EC 1.1.1.2), a glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and a diaphorase (EC 1.6.99.2), wherein the stabilized coenzyme is a compound of general formula (II):
- in which
- A=adenine or an analogue thereof;
- T=in each case independently denotes O or S;
- U=in each case independently denotes OH, SH, BH3−, or BCNH2−;
- V=in each case independently denotes OH or a phosphate group, or two groups which form a cyclic phosphate group;
- W=COOR, CON(R)2, COR, or CSN(R)2 in which R in each case independently denotes H or a C1-C2 alkyl;
- X1, X2=in each case independently denote O, CH2, CHCH3, C(CH3)2, NH, or NCH3;
- Y=NH, S, O, or CH2;
- Z=a linear residue with 4-6 C atoms or a saturated or an unsaturated carbocyclic or heterocyclic 5-membered ring of general formula (III):
- in which a single or double bond may be present between R5′ and R5″, where
- R4=in each case independently denotes H, F, Cl, or CH3;
- R5=CR42;
- R5′=O, S, NH, NC1-C2-alkyl, CR4, CHOH, or CHOCH3, and R5″=CR42, CHOH, or CHOCH3 if there is a single bond between R5′ and R5″;
- R5′=R5″=CR4 if there is a double bond between R5′ and R5″;
- and
- R6, R6′=in each case independently denote CH or CCH3;
- wherein the Z and the pyridine residue are not linked by a glycosidic bond, and
- wherein enzyme activity decreases by less than 50% after the period of storage of at least 2 weeks at a temperature of at least 20° C. when compared to an initial value of enzyme activity.
16 more elements
Dependencies are inferred from the claim text.