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Method for determining enzyme activity

A technology of enzymatic activity and enzyme-cleaved substrates, which is applied in the field of enzyme detection, can solve the problems of large molecular weight of recombinant proteins, time-consuming, complicated testing process, etc., and achieve accurate detection, reduce treatment costs, improve survival rate and quality of life Effect

Inactive Publication Date: 2020-02-07
北京东西分析仪器有限公司
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Problems solved by technology

The molecular weight of the recombinant protein is large, and the testing process is complex and time-consuming. It is difficult to meet the needs of large-scale screening of thrombotic thrombocytopenic purpura, and it is not suitable for the clinical needs of emergency diagnosis and treatment of patients in the emergency room.
[0009] In summary, there are obviously inconveniences and defects in the actual use of the existing technology, so it is necessary to improve

Method used

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  • Method for determining enzyme activity
  • Method for determining enzyme activity
  • Method for determining enzyme activity

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Embodiment Construction

[0040] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention and are not intended to limit the invention.

[0041] see figure 1 , the invention provides a method for measuring enzyme activity, comprising the following steps:

[0042] S1: Recombinant protein purification of enzyme-cleaved substrates of metalloproteases;

[0043]The amino acid fragment and the histidine at the end are inserted into the bacterial expression vector and cloned, then the bacteria are collected and lysed, centrifuged after the lysis is completed, and the supernatant of the centrifuged bacteria is collected, Then add an equal volume of buffer solution to dilute, and use a reducing ion column to elute the diluted ...

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Abstract

The invention is applicable to the technical field of enzyme detection and provides a method for determining enzyme activity. The method comprises the following steps: S1, performing recombinant protein purification on an enzyme digestion substrate of metalloproteinase; S2, performing an enzyme digestion reaction on the enzyme digestion substrate; and S3, reading a metal target plate after the enzyme digestion reaction, and calculating the enzyme digestion activity in a sample. Therefore, the method has the advantages of good sensitivity, good accuracy, simple operation, low price and wide application range.

Description

technical field [0001] The invention relates to the technical field of enzyme detection, in particular to a method for measuring enzyme activity. Background technique [0002] Thrombotic microangiopathy mainly includes four types of diseases: thrombotic thrombocytopenic purpura, hemolytic uremic syndrome, malignant hypertensive renal damage, and pre-eclampsia renal damage. [0003] Clinically, thrombotic microangiopathy mainly manifests as thrombocytopenia, hemolytic anemia, and organ involvement caused by platelet thrombus in the microcirculation. [0004] The disease involves a wide range of clinical departments, and patients may often seek treatment in different departments, such as nephrology, hematology, neurology, obstetrics and gynecology, dermatology, cardiovascular medicine, and respiratory medicine. Therefore, improving the awareness of this type of disease has become a hot spot in the relevant professional fields at home and abroad. [0005] The clinical manifes...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/37
CPCC12Q1/37G01N2333/8146G01N2800/222G01N2800/226G01N2800/347
Inventor 马龙华李宁顾好粮李晓雯高利艳沈鹏飞王晓东孙斌王利刘松坡谢文君邹飞青龙皎张杰
Owner 北京东西分析仪器有限公司
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