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IV type collagenase content measuring method

A method of determination, the technology of collagenase, is applied in the direction of analyzing materials through chemical reactions and observing the influence of chemical indicators, etc., which can solve the problems of complex analysis steps, difficulties in meeting clinical detection requirements, and expensive kits. And other issues

Inactive Publication Date: 2010-09-29
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The analysis time of substrate gel electrophoresis zymography is as long as several days, and the analysis steps are complicated
Although the ELISA method has the advantages of specificity and high sensitivity, the cost of the required labeled antibody is high, the kit is expensive, the measured enzyme content cannot show the biological activity of the enzyme, and the analysis speed is slow
Although HPLC has the advantages of fast analysis speed and accurate quantification, the specificity of the method is poor, and large samples cannot be detected at the same time, which is difficult to meet the clinical detection requirements.

Method used

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  • IV type collagenase content measuring method
  • IV type collagenase content measuring method
  • IV type collagenase content measuring method

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Embodiment 1

[0018] Embodiment 1: In this embodiment, the content of type IV collagenase in a serum sample is determined. Firstly, the blood sample to be tested is processed. The whole blood sample is centrifuged at 2500 rpm for 5 minutes, and the supernatant can be taken for detection, or the sample can be stored at -20°C or -80°C, but repeated freezing and thawing should be avoided. It should be noted that the serum should be diluted 8 times before testing, and it should be stored at 4°C for less than 1 week, and it should be sealed at -20°C or -80°C. Months, specimen hemolysis will affect the final test results, so hemolysis specimens are not suitable for this test.

[0019] Then start to measure the content of type IV collagenase in the serum sample, please prepare all the reagents in advance before the measurement. When diluting reagents or samples, they need to be mixed evenly, and try to avoid foaming when mixing. A standard curve should be prepared for each test. If the sample c...

Embodiment 2

[0022] Embodiment 2: Utilizing the method of the present invention, this embodiment measures the plasma type IV collagenase content of 112 healthy people, and its content ranges from 31.51 to 49.99 mg / L, with an average value of 41.59 mg / L and a standard deviation of 4.19 mg / L. Statistical analysis confirmed that the plasma type IV collagenase content of healthy people obeyed the normal distribution, so by ±1.96 s The 95% cut-off value of both sides was 33.38-49.80mg / L. The distribution of collagenase type IV in the plasma of the 112 healthy people is as follows: figure 2 shown.

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Abstract

The invention discloses an IV type collagenase content measuring method, which relates to a method for measuring the content of IV type collagenase in blood plasma and blood serum of human bodies. The method is characterized in that the IV type collagenase has the gelatin hydrolyzing specific binding site, other matrix metalloproteinase existing in the blood plasma is difficult to hydrolyze the gelatin, TNBS is adopted for developing the color of gelatin hydrolysates, and the quantitative analysis on the specificity of the IV type collagenase in the living blood serum of the blood plasma of the human bodies can be carried out. The invention can accurately and fast measure the IV type collagenase content in the blood plasma or the blood serum of the human bodies.

Description

technical field [0001] The invention relates to a method for measuring the content of type IV collagenase in human plasma and serum. Background technique [0002] Type IV collagenase is an important matrix metalloproteinase in organisms, also known as basement membrane collagenase, which plays an important pathophysiological role in the processes of cell migration, wound repair, invasion and metastasis of malignant tumors. Current studies have shown that the abnormal increase of plasma type IV collagenase content is often closely related to heart failure, emphysema, autoimmune diseases, tumor growth, invasion and metastasis of tumor cells and other major diseases. In order to study the pathophysiological effects of type Ⅳ collagenase more extensively, it is of great significance to establish a fast, accurate, low-cost and high-throughput detection method. [0003] At present, there are three main methods for detecting the activity of plasma type IV collagenase: substrate ge...

Claims

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Application Information

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IPC IPC(8): G01N21/78
Inventor 凌青余谨赵云斌
Owner HUAZHONG UNIV OF SCI & TECH
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