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Determining method for activity of apolipoprotein J

A technology for measuring apolipoproteins and methods, which is applied in the field of determination of apolipoprotein J activity, can solve problems such as not pointing out apolipoproteins, and achieve the effect of easy operation and high accuracy

Pending Publication Date: 2021-08-20
厦门德馨尚品医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is not indicated whether ApoJ also has a protective effect on the enzymatic activity of GST

Method used

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  • Determining method for activity of apolipoprotein J
  • Determining method for activity of apolipoprotein J
  • Determining method for activity of apolipoprotein J

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: Detection of the activity of recombinant human apolipoprotein J

[0050] 1.1 Solution preparation:

[0051] All the above solutions were prepared, bovine GST enzyme and BSA were purchased from sigma company;

[0052] Recombinant apolipoprotein J solvent (GSH buffer) preparation: use 20mM, pH7.4 PB buffer, add 58.8g of sodium citrate dihydrate, 1.54g of GSH and 0.61g of L-Cys, use NaOH / HCL Adjust the pH value to 7.4, and finally use 20mM, pH7.4 PB buffer to make up to 1L volume;

[0053] Recombinant apolipoprotein J is a certain batch of products of our company. The concentration is: 1.53mg / ml.

[0054] 1.2 Thermal aggregation experiment

[0055] 1) Prepare different ratio reaction points according to the experimental requirements, and the sample addition expression is as follows:

[0056] Sample final concentration ratio table (1:1)

[0057] Numbering components GST(μg / ml) ApoJ (μg / ml) BSA (μg / ml) 1 GST alone 187.5 0 0 2...

Embodiment 2

[0076] Embodiment 2: Detection of the activity of recombinant human apolipoprotein J

[0077] Take another batch of apolipoprotein J samples, the protein concentration is 2.2 mg / ml; the samples are set with different concentration ratios according to the same method as in Example 1, and then heat-treated and GST is used to measure the enzyme activity;

[0078] Such as image 3 As shown, according to the GST specific activity as the Y axis, and the different concentrations of apolipoprotein J as the X axis, the GST activity data after heat treatment was used to make an S-shaped curve. Taking 1 / 2 of the initial specific activity of GST before heating as EC50, according to the curve fitting formula, the concentration value on the X-axis corresponding to EC50 is 0.099199 mg / ml;

[0079] According to the definition of activity, when the concentration of apolipoprotein J is 0.099199 mg / ml, the corresponding activity of apolipoprotein J is 1 unit / ml, and the specific activity of the...

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Abstract

The invention provides a determining method for activity of apolipoprotein J. The method comprises the following steps: adding a proper amount of the apolipoprotein J into glutathione S-transferase, and calculating the activity of the apolipoprotein J by determining the change of the specific activity of glutathione S-transferase before and after heat treatment. According to the method, the specific activity of the apolipoprotein J is determined by detecting the activity change of the glutathione S-transferase before and after heating by utilizing the characteristic that the apolipoprotein J can inhibit protein from forming aggregation and precipitation after heating for the first time, the related operation is simple, and the accuracy is high.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a method for measuring the activity of apolipoprotein J. Background technique [0002] Apolipoprotein J (ApoJ, also known as clusterin) is a multifunctional glycoprotein widely present in various tissues and body fluids of the human body. Apolipoprotein J is a heterodimeric protein with a relative molecular mass of 75-80KDa, which can combine with human plasma high-density lipoprotein (HDL) and very high-density lipoprotein (VHDL). Apolipoprotein J has complex functions and is involved in many physiological processes in vivo, including regulation of complement function, inhibition of apoptosis and inflammation, and participation in lipid transport. It has been found that apolipoprotein J is highly expressed in various disease processes , such as neurodegeneration, atherosclerosis, myocardial infarction, tumors, etc. [0003] Molecular chaperones are a class of proteins that b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48G01N33/92G01N21/31
CPCC12Q1/48G01N33/92G01N21/31G01N2333/775G01N2333/91177
Inventor 陈翔杜佩云蔡文凯
Owner 厦门德馨尚品医疗科技有限公司
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