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Method for high-density lipoprotein chip electrophoresis

A high-density lipoprotein and chip technology, applied in separation methods, chemical instruments and methods, material analysis by electromagnetic means, etc., to achieve the effect of high-efficiency separation and detection, rapid separation and analysis capabilities

Inactive Publication Date: 2009-12-30
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Abstract
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Problems solved by technology

However, this method requires different buffer systems and different separation modes to separate HDL subclasses.

Method used

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  • Method for high-density lipoprotein chip electrophoresis
  • Method for high-density lipoprotein chip electrophoresis
  • Method for high-density lipoprotein chip electrophoresis

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Experimental program
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Embodiment Construction

[0017] 1) Selection of electrophoresis buffer system:

[0018] The buffer solution (Background electrolyte) has a pH of 8.5 and consists of 40mmol / L tricine, 50mmol / L meglumine, and 0.02mmol / L sodium dodecyl sulfate.

[0019] 2) Preparation and labeling of serum samples

[0020] After three days of vegetarianism, 3 mL of blood was collected on an empty stomach in the morning, and the serum was separated by centrifugation at 3000 r / min for 10 min. The serum sample was added to the same volume of precipitant with a pH of 6.1 composed of 0.05 mol / L magnesium chloride and 1.52 mmol / L phosphotungstic acid. , mix well, place at room temperature for 15 minutes, and then centrifuge at 3000 r / min for 15 minutes. In the supernatant after centrifugation, only one lipoprotein, HDL, remains for determination. It is required to complete the separation and detection within 4 hours, or store at -70°C. Add 2 μL deionized water to 6 μL serum, then add 4 μL 0.2 g / L NBD C6-ceramide (pre-dissolv...

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Abstract

The invention discloses a method for high-density lipoprotein chip electrophoresis. The method comprises the following steps of selection of a buffer system, selection of an electrophoresis chip, sampling and marking of blood serum, electrophoresis and the like. The method is based on the combination of chip electrophoresis technology and a laser induced fluorescence detection system, and can separate two sub-categories of the blood serum HDL in 4 minutes, and make up the defect that the sub-categories of the HDL cannot be analyzed by a conventional blood biochemical analysis method. Due to the high-resolution and quick separation analysis capability shown by the chip electrophoresis to lipoprotein, the method hopefully becomes one of analysis methods for separating and detecting the sub-categories of the HDL quickly, conveniently and efficiently.

Description

Technical field: [0001] The invention relates to a high-density lipoprotein chip electrophoresis method. Background technique: [0002] Lipoproteins are nanometer-sized particles, which can be divided into three main categories according to ultracentrifugation: high-density lipoprotein (High Density Lipoprotein, HDL), low-density lipoprotein (lowdense lipoprotein, LDL) and very low-density lipoprotein ( Very Low Density Lipoprotein, VLDL). HDL is a lipoprotein composed of lipids and proteins, which has large heterogeneity in shape, density, particle size, charge and physical and chemical properties, and is mainly divided into HDL 2 and HDL 3 Two subclasses. [0003] At present, domestic and foreign reports have analyzed HDL subclasses from multiple aspects, mainly including: density gradient ultracentrifugation (Density Gradient Ultracentrifugation, DGUC), gradient gel electrophoresis (Gradient Gel Electrophoresis, GGE), proton nuclear magnetic resonance (Nuclear Magneti...

Claims

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Application Information

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IPC IPC(8): G01N27/447B01D57/02
Inventor 王惠民郑慧斐金庆辉丛辉
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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