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A composition and its application in detecting the activity of lipoprotein-associated phospholipase a2

A technology of composition and lipoprotein, which is applied in the field of medical testing, can solve the problems of poor stability of reagents, decreased stability of substrates, inconvenient use, etc., and achieve the effect of not being easily hydrolyzed and prolonging the validity period

Active Publication Date: 2021-07-30
SHENZHEN AMTECH BIOENGINEERING LTD INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the substrate PAF analogs of this method are lipids, which are almost insoluble in water, and serum enzymes in the sample can also hydrolyze PAF analogs, making the determination results inaccurate
Although the use of ethanol or glycerin can promote the substrate to form a water phase, making the measurement results accurate, but the stability of the substrate is greatly reduced, and it is easy to degrade, which in turn affects the blank absorbance of the reagent and the validity period of the reagent.
Currently commercially available kits are mostly three reagents of R1, R2a, and R2b. R2a and R2b need to be mixed in proportion before use. The validity period of the mixed reagent is 28 days, which is inconvenient to use. The stability of the reagent after preparation is poor and the validity period is short.

Method used

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  • A composition and its application in detecting the activity of lipoprotein-associated phospholipase a2
  • A composition and its application in detecting the activity of lipoprotein-associated phospholipase a2
  • A composition and its application in detecting the activity of lipoprotein-associated phospholipase a2

Examples

Experimental program
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Embodiment 1

[0065] The detection reagent combination of the present embodiment comprises:

[0066] The composition of reagent 1 (or called R1 reagent, the first reagent) is as follows:

[0067] 100mmol / L 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), 15mmol / L ethylenediaminetetraacetic acid (EDTA), 9mmol / L 3-[3-(cholamidopropyl)dimethylaminopropanesulfonic acid Salt (CHAPS), 10mmol / L sodium n-nonane sulfonate, 200mmol / L sodium chloride, the balance is water; the pH of reagent 1 is 7.6.

[0068] The composition of reagent 2 (or called R2 reagent, second reagent) is as follows:

[0069] 20mmol / L citric acid, 15mmol / L sodium n-nonanesulfonate, 5wt% ethanol, 3mmol / L 1-tetradecanoyl-2-4-p-nitrophenol succinic anhydride-3-phosphatidylcholyl, 0.05wt %18-Crown Ether-6, 0.1wt% Dextran 40 (also known as Dextran-40, Dextran-40, Dextran T-40, Dextran T-40), the balance is water; the pH of reagent 2 is 4.5 . The mass percent concentration mentioned in Reagent 2 refers to the mass percent con...

Embodiment 2

[0071] Lp-PLA in embodiment 2 and embodiment 1 2 The only difference in the assay kit is that the stabilizer in reagent R2 is 0.05wt% 18-crown-6, and the others are the same as those in Example 1, and will not be repeated here.

Embodiment 3

[0073] Embodiment 3 and the Lp-PLA in embodiment 1 2 The only difference in the assay kit is that the stabilizers in the reagent R2 are 0.05wt% 15-crown-5 and 0.1wt% dextran 40. Others are the same as in Example 1 and will not be repeated here.

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Abstract

A composition and its application in detecting the activity of lipoprotein-associated phospholipase A2, the composition contains a substrate and a stabilizer, and the stabilizer is selected from at least one of crown ether compounds and dextran compounds , the substrate can be bound to lipoprotein-associated phospholipase A2, and the substrate bound to the lipoprotein-associated phospholipase A2 will be decomposed. The addition of the stabilizer makes the substrate more stable, less prone to hydrolysis, effectively controls the blank absorbance of the reagent, and makes the validity period of the reagent more than 12 months.

Description

technical field [0001] The invention relates to the technical field of medical testing. In particular, it relates to a composition and its application in detecting the activity of lipoprotein-associated phospholipase A2. Background technique [0002] Studies have shown that the prevalence of cardiovascular disease in China and other countries is on the rise. Studies have shown that atherosclerosis is the pathological basis of cardiovascular and cerebrovascular diseases. [0003] Lipoprotein-associated phospholipase A2 (Lp-PLA for short) 2 ) can generate lysophosphatidylcholine (lysophosphatidylcholine, Lyso-PC) and oxidized free fatty acids (oxidized free fatty acids, ox-FA) by hydrolyzing oxidized phospholipids on the oxidized low-density lipoprotein (LDL) on the arterial intima, These two pro-inflammatory mediators can stimulate the production of adhesion factors and cytokines, promote the occurrence and development of atherosclerosis, and lead to thrombosis and cardiov...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/44
CPCC12Q1/44C12Q2334/10G01N2333/918
Inventor 陈小茹吴传侠吴向东周博
Owner SHENZHEN AMTECH BIOENGINEERING LTD INC
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