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Hexokinase freeze-dried powder preparation method

A technology of hexokinase and freeze-dried powder, which is applied in the field of preparation of hexokinase freeze-dried powder, can solve the problems of less hexokinase and no involvement of hexokinase dry powder, etc., to facilitate preservation and solve the problem of serious loss of enzyme activity Effect

Active Publication Date: 2018-05-18
河北省微生物研究所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are relatively few studies on hexokinase in China. The applicant disclosed the preparation method of hexokinase for diagnosis in the patent document with the publication number CN106479992A. Preparation of hexokinase dry powder

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A preparation method of hexokinase freeze-dried powder consists of the following process steps:

[0026] A. Prepare the hexokinase pure enzyme solution with an enzyme activity of 200U / ml according to the method in the publication number CN106479992A;

[0027] B. Add 3% (m / v) sucrose, 1% (m / v) lactose, and 6% (m / v) sorbitol to the hexokinase pure enzyme solution prepared in step A as a freeze-dried protective agent, and place Store at 4°C for 4-5h to make the protective agent fully dissolved and evenly distributed in the enzyme solution;

[0028] C. Pre-freeze for 12-14 hours at a pre-freezing temperature of -20°C, and freeze-dry for 16-20 hours in a vacuum freeze-drying agent to obtain white hexokinase freeze-dried powder, which is stored at -20°C.

Embodiment 2

[0030] The difference between this embodiment and embodiment 1 is:

[0031] A preparation method of hexokinase freeze-dried powder consists of the following process steps:

[0032] A. Prepare the hexokinase pure enzyme solution with an enzyme activity of 300U / ml according to the method in the publication number CN106479992A;

[0033] B. Add 5% (m / v) sucrose, 2% (m / v) lactose, and 8% (m / v) sorbitol to the pure enzyme solution of hexokinase prepared in step A as a freeze-dried protective agent. Store at 4°C for 4-5h to make the protective agent fully dissolved and evenly distributed in the enzyme solution;

[0034] The rest is the same as in Example 1.

Embodiment 3

[0036] The difference between this embodiment and embodiment 1 is:

[0037] A preparation method of hexokinase freeze-dried powder consists of the following process steps:

[0038] A. Prepare the hexokinase pure enzyme solution with an enzyme activity of 250 U / ml according to the method in the publication number CN106479992A;

[0039] B. Add 4% (m / v) sucrose, 1.5% (m / v) lactose, and 7% (m / v) sorbitol to the pure enzyme solution of hexokinase prepared in step A as a freeze-dried protective agent. Store at 4°C for 4-5h to make the protective agent fully dissolved and evenly distributed in the enzyme solution.

[0040] The rest is the same as in Example 1.

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PUM

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Abstract

The invention belongs to preparation of enzyme preparations and especially relates to a hexokinase freeze-dried powder preparation method. According to the preparation method, hexokinase pure enzyme fluid is prepared according to a method with a publication number as CN106479992A; saccharose, lactose and sorbitol are added into the hexokinase pure enzyme fluid to serve as a freeze-drying protecting agent, and the protecting agent is fully dissolved in the enzyme fluid to be evenly distributed; after pre-freezing, the hexokinase pure enzyme fluid is put in a vacuum freeze-drying agent to be frozen and dried to obtain white hexokinase freeze-dried powder. The hexokinase freeze-dried powder preparation method disclosed by the invention solves the problem that an inactivation rate is high whenthe hexokinase is directly frozen and dried at present; the hexokinase freeze-dried powder preparation method has the advantages that a hexokinase freeze-dried powder enzyme activity preserving rateis high, enzyme property of the hexokinase freeze-dried powder is stabler relative to liquid-state hexokinase, the hexokinase freeze-dried powder can meet a use requirement in national standards to kits, and the hexokinase freeze-dried powder is more favorable for being stored and the like.

Description

Technical field [0001] The invention belongs to the preparation of enzyme preparations, in particular to a preparation method of hexokinase freeze-dried powder. Background technique [0002] Serum glucose (GLU) testing is widely used in the diagnosis of diabetes, observation of curative effects and general clinical diagnosis. The hexokinase method has a strong ability to act on α-D-glucose due to its few interference factors, a small Km value, and a strong affinity for the substrate. It has a short measurement time when prepared into a finished kit, and is especially suitable for automatic analyzers. It is especially suitable for emergency testing and is recognized as a reference method for determining serum glucose. [0003] In the process of production and use, hexokinase needs to be stored for a long time. The storage time is short at 4°C in liquid state. Frozen storage at -20°C will cause aggregation, modification, degradation, and pollution due to repeated freezing and thawin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/96C12N9/12
CPCC12N9/1205C12N9/96C12Y207/01001
Inventor 刘春卯曹倩荣郑翔秦艳梅马清河
Owner 河北省微生物研究所有限公司