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Method for long-term stabilization of inositol dehydrogenase, ketoamine oxidase and sphingomyelinase in liquid

A technology of ketamine oxidase and alcohol dehydrogenase, which is applied in the field of in vitro diagnostic reagents, can solve the problems of low enzyme activity, influence on patient diagnosis and treatment plan, poor stability of enzyme protein, etc.

Active Publication Date: 2019-06-04
深圳市安帝宝科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in such biochemical diagnostic reagents, enzymes exist in liquid reagents in the form of solutes. In practice, it is found that enzyme proteins stored in solutions are sometimes less stable.
Even if the transportation and storage environment of the reagents are strictly controlled, the activity of the enzymes in the reagents may decline as the storage time of the reagents prolongs, which may affect the accuracy of the reagents during use
Therefore, the stability of enzymes in liquid is directly related to the quality of biochemical diagnostic reagents, which may affect doctors' diagnosis and treatment plans for patients.

Method used

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  • Method for long-term stabilization of inositol dehydrogenase, ketoamine oxidase and sphingomyelinase in liquid
  • Method for long-term stabilization of inositol dehydrogenase, ketoamine oxidase and sphingomyelinase in liquid
  • Method for long-term stabilization of inositol dehydrogenase, ketoamine oxidase and sphingomyelinase in liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Long-term stability test of inositol dehydrogenase

[0020] Prepare liquid reagent A containing inositol dehydrogenase and stabilizer as follows:

[0021] [1] Weigh 23.73g of 3-(cyclohexylamine)-2-hydroxypropanesulfonic acid and 9g of sodium chloride and dissolve them in 975ml of deionized water;

[0022] [2] Add 1.5 g of benzyl dimethylphenol polyoxyethylene ether and 0.1 g of 4-formylphenylboronic acid to the solution in [1] in sequence, and adjust the pH to 9.0 with 0.1M HCl, and finally set the volume to 1 L;

[0023] [3] Add 1000 mg of inositol dehydrogenase to the solution in [2], and finally configure the inositol dehydrogenase liquid reagent.

[0024] Prepare the control liquid reagent B containing inositol dehydrogenase without stabilizer as follows:

[0025] [1] Weigh 23.73g of 3-(cyclohexylamine)-2-hydroxypropanesulfonic acid and 9g of sodium chloride and dissolve them in 975ml of deionized water;

[0026] [2] Adjust the pH of the solution in ...

Embodiment 2

[0035] Embodiment 2: Long-term stability test of ketoamine oxidase

[0036] Prepare liquid reagent C containing ketoamine oxidase and stabilizer as follows:

[0037] [1] Weigh 13.6g of potassium dihydrogen phosphate and 9g of sodium chloride and dissolve them in 975ml of deionized water;

[0038] [2] Add 1.5g of benzyldimethylphenol polyoxyethylene ether and 0.1g of 4-formylphenylboronic acid to the solution in [1] in sequence, and adjust the pH to 7.5 with 1M sodium hydroxide, and finally set the volume to 1L ;

[0039] [3] Add 1000 mg of ketoamine oxidase to the solution in [2], and finally configure the liquid reagent of inositol dehydrogenase.

[0040] Prepare the control liquid reagent D containing ketoamine oxidase without stabilizer as follows:

[0041] [1] Weigh 13.6g of potassium dihydrogen phosphate and 9g of sodium chloride and dissolve them in 975ml of deionized water;

[0042] [2] Adjust the pH of the solution [1] to 7.5 with 0.1M sodium hydroxide, add 1000 ...

Embodiment 3

[0051] Embodiment 3: Long-term stability test of sphingomyelinase

[0052] Prepare liquid reagent C containing sphingomyelinase and stabilizer as follows:

[0053] [1] Dissolve 15.7g of tris(hydroxymethyl)aminomethane and 9g of sodium chloride in 975ml of deionized water;

[0054] [2] Add 1.5g of benzyldimethylphenol polyoxyethylene ether and 0.1g of 4-formylphenylboronic acid to the solution in [1] in sequence, and adjust the pH to 8.0 with 1M HCl, and finally set the volume to 1L;

[0055] [3] Add 1000 mg of sphingomyelinase to the solution in [2], and finally make sphingomyelinase liquid reagent E.

[0056] Prepare the control liquid reagent F containing sphingomyelinase without stabilizer as follows:

[0057] [1] Dissolve 15.7g of tris(hydroxymethyl)aminomethane and 9g of sodium chloride in 975ml of deionized water;

[0058] [2] After adjusting the pH of the solution [1] to 8.0 with 1M HCl, add 1000 mg of sphingomyelinase and finally adjust the volume to 1L, and final...

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PUM

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Abstract

The present invention provides a method for long-term stabilization of inositol dehydrogenase, ketoamine oxidase and sphingomyelinase in liquid. The method is used to prepare an enzymatic biochemicalkit for determining inositol, glycated albumin and small and dense low density lipoprotein cholesterol for a purpose of enabling the enzymatic biochemical kit to have a shelf life of 12 months or more. The method is characterized in that a composition of 4-formylbenzeneboronic acid and benzyldimethylphenol polyoxyethylene ether is added to a reagent solution for preserving enzymes, and concentrations of the 4-formylbenzeneboronic acid and benzyldimethylphenol polyoxyethylene ether are preferably 0.008-0.012 w / V% and 0.1-0.2 w / V%, respectively.

Description

technical field [0001] The invention relates to a method for stabilizing inositol dehydrogenase, ketoamine oxidase and sphingomyelinase in liquid for a long time, which is used for preparing high-stability in vitro diagnostic reagents. Background technique [0002] In Vitro Diagnosis, namely In Vitro Diagnosis (IVD), refers to the acquisition of clinical diagnostic information by testing human samples (blood, body fluids, tissues, etc.) outside the human body. At present, in vitro diagnostic reagents are an indispensable means for doctors to diagnose and treat, and play an important role in the diagnosis, treatment, curative effect monitoring, prevention and prognosis judgment of patients. [0003] Biochemical reagents using enzymatic methods are the main category of in vitro diagnostic reagents, and have the advantages of simple operation, convenience, and low cost. Such in vitro diagnostic reagents are usually prepared as two-liquid reagent kits, and the detection process...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/96C12N9/02C12N9/04C12N9/16
Inventor 蔡泽浪张伯平田金艳杨丽竹谢芳
Owner 深圳市安帝宝科技有限公司
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