A gel electrophoresis system, preparation method and method for separating and detecting low-density lipoprotein using the same

Abstract

The invention provides a gel electrophoresis system, a preparation method and a method for separating and detecting low density lipoprotein by the gel electrophoresis system. The gel electrophoresis system has the capability of separating and detecting various subtypes of plasma LDL (low-density lipoprotein) by adjusting the content of polyacrylamide in a gel electrophoresis separation column andtypes of electrophoresis buffers, and has the following advantages: (1) the separation effect and resolution are far superior to those of existing products, and the electrophoresis operation procedureand separation time are further shortened; (2) electrophoresis separation and detection can be realized only by 2 layers of gel, while existing kit products need at least 3 layers of gel; (3) the cost is low, the production cost is lower than 6 yuan, and is far lower than 120 yuan of existing Lipoprint LDL Subfragmentations Kit, but detection and separation effects are the same or even better.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a gel electrophoresis system, a preparation method and a method for separating and detecting low-density lipoprotein using the same. Background technique [0002] Cardiovascular disease is a disease that seriously threatens human beings. Many factors, such as hypertension, dyslipidemia, diabetes, family history, gender and age, smoking, obesity, etc., can lead to the occurrence of the disease. "Guidelines for the Prevention of Dyslipidemia in Chinese Adults (Revised 2016)" and the "Guidelines for the Management of Dyslipidemia" issued by the European Society of Cardiology and the European Atherosclerosis Society jointly pointed out that LDL cholesterol (LowDensity Lipoproteins -cholesterol (LDL) content belongs to a class of factors leading to dyslipidemia, and it is recommended to reduce the level of LDL in plasma as the most important intervention target for bloo...

AI Technical Summary

Problems solved by technology

The original polypropylene gel electrophoresis method was proposed by Ornstein, Davis and Leammli et al. It is widely used in the biochemical and pharmaceutical analysis of proteins. However, the traditional gel system has a low resolution for plasma lipoproteins. Most of the gel systems are alkaline systems with high pH, ​​which will lead to continuous hydrolysis of polyacrylamide gels. Even if they are stored at 2°C~8°C for at most 2 weeks, the traditional gel system will lose the separation effect, and the hydrolysis will be shortened. The migration distance of proteins reduces the resolution of protein bands. In order to ensure the experimental results, most laboratories use ready-made gels, which greatly restricts work efficiency and consumes a lot of time and manpower. Moreover, the market Most of the precast gels above cannot be used for the fine separation of lipoprotein subtypes

[0006] In the prior art, the gel electrophoresis system commonly used to separate lipoproteins in plasma, such as the gel electrophoresis system disclosed in CN1699406A, includes separation gels 1 to 3 with polyacrylamide content of 3 wt%, 3.6 wt% and 7 wt% respectively , which is difficult to separate the subtypes of LDL in plasma

At present, the use of gel electrophoresis to separate the subtypes of lipoproteins in plasma is relatively mature. The Lipoprint LDL Subfractions Kit developed and produced by Quantimetrix Company of the United States can not only refine and classify LDL into 7 subcomponents and Detect the expression level of each subcomponent, and simultaneously detect the levels of CM (chylogranules), VLDL (very low-density lipoprotein), LDL and HDL (high-density lipoprotein) and the proportion of lipoprotein components in total cholesterol Bi, is currently the only product approved by the FDA for the detection and analysis of low-density lipoprotein subtypes. However, the unit price of this product is about 120 yuan, and the import cost is very high

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