Preparation method of high purity and low density lipoprotein

A low-density lipoprotein, high-purity technology, applied in the field of material extraction, can solve the problems of less conventional extraction, affecting dialysis efficiency, and difficulty in meeting the requirements of blood collection, and achieve the effect of improving the recovery rate.

Inactive Publication Date: 2019-02-12
XUZHOU CENT HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, many laboratory dialysis bags are recycled after cleaning, sterilization and other steps. Repeated treatment of dialysis bags will significantly affect the efficiency of dialysis.
The amount of routine extraction of plasma LDL is generally small. For example, when a large amount of LDL is required in the experiment, the amount of blood colle...

Method used

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  • Preparation method of high purity and low density lipoprotein
  • Preparation method of high purity and low density lipoprotein
  • Preparation method of high purity and low density lipoprotein

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Experimental program
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Embodiment Construction

[0028] The present invention will be further described below based on the drawings and embodiments.

[0029] 1. Plasma separation:

[0030] Ethylenediaminetetraacetic acid (EDTA) is added to the plasma, the final concentration is 1mmol / L, 3000rpm×20min, the precipitation is blood cells, and the supernatant plasma is ready for use.

[0031] 2. CM separation:

[0032] Centrifuge: Beckman Avanti J-E high-speed refrigerated centrifuge;

[0033] Rotor, centrifuge tube: JA-20: 20000rpm, 18400×g, 8×50ml;

[0034] Centrifugation parameters: 4℃, 20000rpm×2h;

[0035] Centrifugation result: CM is white and floats on the surface of the liquid, or close to the wall of the centrifuge tube, carefully suck out.

[0036] 3. Separation of plasma lipoproteins:

[0037] Centrifuge: Hitachi CP-80WX ultra-speed centrifuge;

[0038] Rotor, centrifuge tube: P70AT: 70000rpm, 505000×g, 8×40ml, 40PA tube.

[0039] 3.1, VLDL separation:

[0040] Plasma treatment: mix the plasma after removing CM, add ultrapure water or ...

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Abstract

The invention belongs to the technical field of material extraction, and particularly relates to a preparation method of a high purity and low density lipoprotein. Chylomicrons and very low density lipoproteins in blood plasma are centrifuged and removed according to different densities, remain blood plasma is extracted and added withpotassium bromide and the density is adjusted to 1.045g/mL, centrifugation is carried out again to obtain a normal low density lipoprotein located at the uppermost layer, and a phosphate buffer salt solution is used for dialyzing to reduce K+ concentration; and aresultof the K+ concentration of the low density lipoprotein before and after dialyzing measured by a full-automatic biochemical analyzer is compared with a result measured by an osmotic pressure analyzer, if the K+ concentration of the low density lipoprotein is dialyzed to be with no obvious differences with the osmotic pressure of the phosphate buffer salt solution, the potassium bromide is considered to be completely dialyzed, and then the high purity and low density lipoprotein is obtained. The preparation method of the high purity and low density lipoprotein can obtain the high purity and low density lipoprotein to meet of existing experimentalrequirements.

Description

Technical field [0001] The invention belongs to the technical field of material extraction, and specifically relates to a method for preparing high-purity low-density lipoprotein. Background technique [0002] Blood lipids are the general term for cholesterol, triglycerides and lipids in serum. Blood lipids are insoluble in water and must be combined with apolipoprotein to form lipoproteins before they can be dissolved in the blood and transported to tissues for metabolism. Low-density lipoprotein cholesterol (Low-density Lipoprotein Cholesterol, LDL-C) is the lipoprotein with the highest cholesterol content (about 50%) in the blood, so the LDL-C concentration can basically reflect the total amount of blood low-density lipoprotein (LDL). In addition, LDL can be oxidized to oxidized LDL (Oxidized Low-density Lipoprotein, Ox-LDL). [0003] Three common ultracentrifugation methods used for lipoprotein analysis are: analytical ultracentrifugation, zone ultracentrifugation, and prepara...

Claims

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Application Information

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IPC IPC(8): G01N1/34G01N33/68
CPCG01N1/34G01N33/6812
Inventor 庞慧
Owner XUZHOU CENT HOSPITAL
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