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Electrophoretic Titration Method for Determination of Peroxidase Activity

A technology of peroxidase and activity, which is applied in the field of bioanalysis technology and medical inspection, and can solve the problems of large sample consumption, high cost, and low throughput

Active Publication Date: 2019-02-22
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods have many advantages, but at the same time face some shortcomings, such as low throughput, high cost and large sample consumption, etc.

Method used

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  • Electrophoretic Titration Method for Determination of Peroxidase Activity
  • Electrophoretic Titration Method for Determination of Peroxidase Activity
  • Electrophoretic Titration Method for Determination of Peroxidase Activity

Examples

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Embodiment 1

[0079] In this example, the horseradish peroxidase activity was measured by electrophoresis titration method, and the electrophoresis titration method is as figure 1 Shown. First fill the channel part of the chip with gel, inject the mixed solution a of hydrochloric acid solution and background electrolyte solution into the anode pool, and inject the mixed solution b of the chemiluminescent base solution and the background electrolyte solution into the cathode pool; then immediately pass the horseradish The oxidase solution is injected into the cathode cell to make it catalyze the chemiluminescence substrate reaction to generate blue fluorescent excited luminol anion L*; after the enzyme catalyzed reaction is stable for a period of time, apply a voltage to make the excitation generated in the cathode cell The state luminol anion L* moves toward the anode and matches the hydrogen ion H moving toward the cathode in the anode pool. + The MRB is formed, and the MRB moves to the ano...

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Abstract

The present invention discloses an electrophoresis titration method for determining the activity of peroxidase. The electrophoresis titration method comprises: filling the channel part of a chip with a gel, injecting the mixed solution a of a hydrochloric acid solution and a background electrolyte solution into an anode pool, and injecting the mixed solution b of a chemiluminescent base solution and a background electrolyte solution into a cathode pool; injecting a peroxidase solution into the cathode pool to catalyze the chemiluminescent base solution to react so as to produce excitation state luminol anions L*; after the enzyme catalytic reaction is stabilized, applying an electric field to make the excitation state luminol anions L* generated in the cathode pool and the hydrogen ions H<+> in the anode pool form MRB, wherein the generated MRB moves to the anode; and according to the functional relationship between the MRB movement rate and the activity of the peroxidase, determining the activity index of the peroxidase. With the method of the present invention, various enzyme indexes of the peroxidase can be effectively and rapidly determined without complex optical detection equipment.

Description

Technical field [0001] The invention relates to the fields of biological analysis technology and medical testing, and specifically relates to an electrophoresis titration method for measuring peroxidase activity. Background technique [0002] Enzyme analysis is used in clinical medicine (Abraham, EC; Huff, TA; Cope, ND; Wilson, JB; Bramsome ELISA, EDJr.; Huisman, THJDiabetes 1978, 27, 931-937.) Biology (Tannu, NS; Hemby ,SENature Protocols 2006,1,1732-1942.) and drug screening (Li,Juan; Wu,Ling-Jie; Guo,Shan-Shan; Fu,Hua-E; Chen,Guo-Nan; Yang,Huang-Hao ,NANOSCALE,2013,5,619-623.) has an important role in the field. Traditional detection methods are generally based on the detection of substrate reduction and product generation in the enzyme reaction system to determine the catalytic rate, activity, and Michaelis constant. There are many methods that can be used for enzyme detection, such as mass spectrometry (Northen, TR; Lee, JC; Hoang, L; Raymond, J; Hwang, DR; Yannone, SM; Wo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447
CPCG01N27/447
Inventor 曹成喜钟冉谢海洋张强沙敏·贾汉肖华樊柳荫
Owner SHANGHAI JIAOTONG UNIV
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