High-density lipoprotein subtyping detection kit and preparation method and application thereof

A technology of high-density lipoprotein and detection kits, which is applied in the direction of measuring devices, electrophoresis analysis, and analysis materials, can solve the problems of cumbersome gradient gel production, difficult import, unfavorable commercial production, etc., and achieve electrophoresis operation procedures and The separation time is shortened, the detection and separation effects are the same, and the effect is suitable for large-scale production

Pending Publication Date: 2021-02-02
SHANGHAI BIOTECAN PHARMA +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, in gradient gel electrophoresis, HDL can be divided into five subclasses according to particle size: HDL2b, HDL2a, HDL3a, HDL3b, and HDL3c. However, the preparation of gradient gels is cumbersome, and the differences between gel batches are large, and sample detection takes a long time. It is long, generally takes more than 24 hours, and the specificity is poor, and the quantitative analysis of HDL subclasses cannot be carried out, so it is still limited to the stage of laboratory research
Another example is the method disclosed in CN105137070A, which includes separation gel 1-3 with polyacrylamide content of 3wt%, 3.6wt% and 7wt%, although the separation of HDL subtypes can be realized, the subtype band separation effect is relatively low. Poor, the resolution is not high, and its gel buffer system is alkaline, which can only be stored for a few days, which is not conducive to commercial production
At present, the relatively mature product for the separation and analysis of HDL subtypes by gel electrophoresis is the Lipoprint HDL Subfractions Kit developed and produced by Quantimetrix in the United States, which includes 3 layers of gel, which can not only subdivide HDL into There are 3 types of large, medium and small, and 10 zones. It can also detect the proportion of HDL subcomponents in HDL simultaneously. However, the import of this product is difficult and costly, and due to the Sino-US trade war , the product has ceased to be supplied to China, and the product and technology are seriously restricted by others

Method used

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  • High-density lipoprotein subtyping detection kit and preparation method and application thereof
  • High-density lipoprotein subtyping detection kit and preparation method and application thereof
  • High-density lipoprotein subtyping detection kit and preparation method and application thereof

Examples

Experimental program
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Embodiment 1

[0098] This embodiment provides a high-density lipoprotein subtyping detection kit, including gel electrophoresis separation column, protection solution, high-density lipoprotein electrophoresis staining solution and electrophoresis buffer powder. Prepared by the following method:

[0099] Prepare the following solutions separately for use in preparing gel columns:

[0100] Protective solution mother solution: Dissolve 30.3g Tris and 10g sucrose in distilled water, add 500μL Proclin300, adjust its pH to 6.7 with HCl, then set the volume to 100mL, filter the above solution with a filter membrane with a pore size of 0.45μm, and store at 4°C Keep away from light.

[0101] Gel mother solution: Dissolve 29g of acrylamide and 1g of methylene bisacrylamide in distilled water, make up to 100mL, filter the above solution with a filter membrane with a pore size of 0.45μm, and store at 4°C in the dark.

[0102] Ammonium persulfate solution: Dissolve 1 g of ammonium persulfate in distil...

Embodiment 2

[0120] This embodiment provides a high-density lipoprotein subtyping detection kit, including gel electrophoresis separation column, protection solution, high-density lipoprotein electrophoresis staining solution and electrophoresis buffer powder. The difference between its preparation method and Example 1 is that in step (2), by changing the addition of the gel mother liquor and distilled water, the content of polyacrylamide in the gel column of the second electrophoresis separation section obtained is 7.0wt %, all other conditions remain unchanged.

Embodiment 3

[0122] This embodiment provides a high-density lipoprotein subtyping detection kit, including gel electrophoresis separation column, protection solution, high-density lipoprotein electrophoresis staining solution and electrophoresis buffer powder. The difference between its preparation method and Example 1 is that in step (2), by changing the addition of the gel mother liquor and distilled water, the content of polyacrylamide in the gel column of the second electrophoresis separation section obtained is 10.0wt %, all other conditions remain unchanged.

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Abstract

The invention relates to a high-density lipoprotein subtyping detection kit and a preparation method and application thereof. The high-density lipoprotein subtyping detection kit comprises a gel electrophoresis separation column, a protective solution, a high-density lipoprotein electrophoresis staining solution and electrophoresis buffer powder, and the gel electrophoresis separation column is immersed in the protective solution. The high-density lipoprotein subtyping detection kit has the capability of separating and detecting each subtyping of plasma lipoprotein HDL, electrophoretic separation and detection can be realized only by two layers of gel, and an existing kit product needs at least three layers of gel; the separation effect and the resolution ratio are far superior to those ofthe existing product, and the electrophoresis operation procedure and the separation time are further shortened; and cost is low, and large-scale production is facilitated.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a high-density lipoprotein subtyping detection kit and a preparation method and application thereof. Background technique [0002] Cardiovascular disease (CVD) is a disease that seriously threatens human beings. Many factors such as hypertension, dyslipidemia, diabetes, family history, gender and age, smoking, obesity and other factors can lead to the occurrence of cardiovascular disease. Among them, dyslipidemia is the trigger An important risk factor for cardiovascular disease and death. [0003] High-density lipoprotein (HDL) is a kind of lipoprotein with extremely heterogeneous composition, density, particle size and function, with a density between 1.063-1.210g / mL. According to the size, density and surface charge of HDL particles, HDL can be divided into subclasses such as HDL2b, HDL2a, HDL3a, HDL3b, and HDL3c. HDL can mediate the reverse cholesterol transp...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): G01N27/447G01N33/487
CPCG01N27/44778G01N33/48707G01N2550/00
Inventor刘亚王政王博伟张宇婷叶琳珊程雅文楼敬伟
OwnerSHANGHAI BIOTECAN PHARMA