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Method for gel electrophoresis separation of serum lipoprotein and quantization detection thereof

A technology of serum lipoprotein and gel electrophoresis, which is applied to the preparation methods of apolipoprotein and peptide, chemical instruments and methods, etc., can solve the problems of weak synchronous expression ability, inconsistent and inhomogeneous reference system in the analysis process, etc. Achieve the effect of realizing metabolic dynamic balance, good application prospect and easy operation

Inactive Publication Date: 2005-11-23
陈允钦 +1
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  • Abstract
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Problems solved by technology

There are three main problems: 1. Due to the great difference in the particle characteristics of different types of serum lipoproteins, it is difficult to take into account the experimental conditions of a single method, and the ability to express them simultaneously is weak. Usually, separate methods are used for detection
2. Due to the inconsistency of the reference system in the analysis process, the results of separate determinations cannot be placed in a unified whole of serum lipoproteins to compare their interrelationships
3. Due to the high requirements of experimental conditions, such as a stable linear concentration gradient, it is difficult to meet the needs of clinical testing
The 2001 survey found that 94.9% of patients with coronary heart disease and hyperlipidemia in my country failed to obtain ideal blood lipid control
Therefore, theoretically, it is difficult to explain the pathogenesis of patients from isolated blood lipid factors
There are some problems as follows: (1) The integrity of the serum lipoprotein particle structure is ignored, and the serum lipoprotein exerts its physiological function under the premise of maintaining the complete particle structure
(2) Ignoring the integrity of serum lipoprotein metabolism, the composition of serum lipoprotein is complex and uneven, and its metabolism is usually in a coordinated and balanced state
(2) The main problems existing in the previous methodology are: (1) The separation methods are mutually independent, and it is difficult to carry out synchronous separation
Due to the great difference in particle characteristics of different types of serum lipoproteins, it is difficult to set electrophoresis conditions and the simultaneous expression ability is poor
(2) There is no unified reference for the analysis method, and a unified comparison cannot be carried out
Due to the neglect of the experimental principle of "uniqueness of the reference system", the measurement results lack mutual comparability, and the overall status of serum lipoprotein metabolic balance cannot be uniformly evaluated

Method used

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  • Method for gel electrophoresis separation of serum lipoprotein and quantization detection thereof
  • Method for gel electrophoresis separation of serum lipoprotein and quantization detection thereof
  • Method for gel electrophoresis separation of serum lipoprotein and quantization detection thereof

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Embodiment Construction

[0015] 1. Reagents

[0016] 1. Storage solution A: 9.60 g of acrylamide (Acr) and 0.25 g of methylene bisacrylamide (Bis), dissolved in deionized distilled water (DDI H 2 O) to 100ml, filter and store in a brown bottle at 4°C for 3 months.

[0017] 2. Storage solution B: Tris (Tris) 18.3g and 1.0M hydrochloric acid 24ml, dissolved in DDI H 2 O to 100ml, pH8.8, filtered and stored at 4°C in a brown bottle.

[0018] 3. Stock solution C: Acr 19.6g and Bis 0.4g, dissolved in DDI H 2 O to 100ml, filter and store in a brown bottle at 4°C for 3 months.

[0019] 4. Stock solution D: Tris 6.06g and disodium edetate (EDTA-Na 2 ) 1.17g, dissolved in DDI H 2 O to 100ml, pH8.8, filtered and stored at 4°C in a brown bottle.

[0020] 5. Initiator: Ammonium persulfate (APS) 1.0g dissolved in DDI H 2 O to 10ml, use instant preparation.

[0021] 6. Staining solution: Dissolve 0.25g of Sudan Black (SB-B) in 25ml of isopropanol, bath in water at 37°C overnight, filter while hot, and store...

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Abstract

The invention discloses a method for gel electrophoresis separation of serum, wherein sectionalized polyacrylamide gels with three different concentrations are used as electrophoresis carrier for forming temperature gradient in the electrophoresis tank, so as to achieve synchronous electrophoretic separation for serum lipoprotein and its subcomponents, wherein the concentrations of the polyacrylamide gel being separation gel 1 3.0%, separation gel 2 3.6%, separation gel 3 7.0%.

Description

technical field [0001] The invention discloses a new method for gel electrophoresis separation of serum lipoproteins and its quantitative detection, relates to a practical method for effectively expressing the metabolic equilibrium state of serum lipoproteins, and is widely used in medical detection and research. Background technique [0002] Serum lipoprotein has a special particle structure, and its composition is complex and heterogeneous. During the metabolic process, it continuously evolves and exchanges substances in sequence, and is usually in a coordinated dynamic equilibrium state. Dyslipoproteinemia is closely related to cardiovascular and cerebrovascular diseases, and it is very important in clinical practice. Over the years, the methods for detecting serum lipoproteins have been continuously improved, such as gradient gel electrophoresis, but there has been a lack of practical methods to effectively express the metabolic balance of serum lipoproteins in clinical ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/26C07K14/775G01N27/447
CPCC07K14/775G01N27/447C07K1/26
Inventor 陈允钦尹恝
Owner 陈允钦
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