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Clinical diagnostic reagent comprising glucose 6-phosphate dehydrogenase (G6PDH), method for stabilizing G6PDH, and use of a stabilizer for g6pdh

A technology of phosphate dehydrogenase and glucose, which is applied in the fields of enzyme stabilization, biochemical equipment and methods, measurement/inspection of microorganisms, etc. It can solve the problems of G6PDH instability and achieve excellent storage stability

Active Publication Date: 2007-01-10
SYSMEX CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the G6PDH in the reagent is unstable, the reagent must contain a G6PDH stabilizer

Method used

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  • Clinical diagnostic reagent comprising glucose 6-phosphate dehydrogenase (G6PDH), method for stabilizing G6PDH, and use of a stabilizer for g6pdh
  • Clinical diagnostic reagent comprising glucose 6-phosphate dehydrogenase (G6PDH), method for stabilizing G6PDH, and use of a stabilizer for g6pdh
  • Clinical diagnostic reagent comprising glucose 6-phosphate dehydrogenase (G6PDH), method for stabilizing G6PDH, and use of a stabilizer for g6pdh

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046]

[0047] The compounds shown below were mixed to prepare a reagent for measuring CK activity. The concentration of each component represents the concentration in the reagent.

[0048] Imidazole 125mM

[0049] EDTA 2.5mM

[0050] Magnesium acetate 12.5mM

[0051] ADP 2.5mM

[0052] AMP 6.25mM

[0053] AP5A 12.5 μM

[0054] Glucose 25mM

[0055] NADP 2.5mM

[0056] NAC 25mM

[0057] G6PDH 1875U / L

[0058] Hexokinase 3750U / L

[0059] In addition, the pH of the reagents was adjusted to 6.6.

[0060] The CK activity assay reagent consists of the above-mentioned reagent (first reagent) and a reagent containing creatine phosphate (second reagent). Since the stability of G6PDH and NAC was analyzed by applying a temperature load to the above-mentioned reagent in this example, it was not used. Second reagent.

[0061] Aminoguanidine was added to the above components to prepare three reagents with different concentrations of aminoguanidine. The concentrations of ami...

Embodiment 2

[0076] Aminoguanidine was added to the reagent of Example 1 at a concentration of 20 mM, and a temperature load was applied at 37° C. for 30 days to measure the activity of G6PDH and quantify the SH group of NAC. In the same manner as in Example 1, activity measurement and quantification were carried out before the temperature load, 7 days after the temperature load, 14 days after the temperature load, 20 days after the temperature load, and 30 days after the temperature load.

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PUM

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Abstract

A clinical diagnostic reagent is described. The reagent comprises glucose 6-phosphate dehydrogenase (G6PDH) and a stabilizer for the G6PDH represented by formula (I): wherein R1 represents a hydrogen, a halogen, a hydroxyl group, a substituted or unsubstituted lower alkyl group, a cycloalkyl group, a lower alkoxy group, a lower alkenyl group, a substituted or unsubstituted aralkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heterocyclic group, a lower alkanoyl group, a substituted or unsubstituted aroyl group, a pyridylcarbonyl group, a lower alkoxycarbonyl group, or a substituted or unsubstituted amino group.

Description

technical field [0001] The invention relates to a clinical diagnostic reagent containing glucose-6-phosphate dehydrogenase (G6PDH), a method for stabilizing G6PDH and the application of the G6PDH stabilizer. Background technique [0002] G6PDH catalyzes the oxidation of glucose-6-phosphate (hereinafter referred to as G6P) to generate 6-phosphate in the presence of coenzymes such as nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP). An enzyme for the reaction of glucono-delta-lactone and reduced NAD (NADH) or reduced NADP (NADPH) or its reverse reaction. Utilizing this catalytic effect, G6PDH can be widely used in various clinical diagnostic reagents. However, since the G6PDH in the reagent is unstable, the reagent must contain a G6PDH stabilizer. [0003] As a reagent containing G6PDH and the aforementioned stabilizer, for example, a reagent for measuring creatine kinase (hereinafter referred to as CK) activity described in USP54...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52G01N33/50G01N21/64C12Q1/32C12N9/96C12N9/04
CPCC12Q1/485C12Q1/32
Inventor 酒井康裕
Owner SYSMEX CORP
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