Kit for detecting concentration of oxidized low-density lipoprotein in sample and preparation method thereof

A low-density lipoprotein and kit technology, which is applied to a kit for detecting the concentration of oxidized low-density lipoprotein in a sample and the field of its preparation, can solve the problems of high detection cost, cumbersome operation, and high skill requirements for experimental operators, and achieves a high level of improvement. Reliability and accuracy, low detection cost, favorable effect of promotion

Inactive Publication Date: 2015-09-16
北京协和洛克生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there are few types of reagents commonly used to detect OxLDL concentration on the market, the detection cost is high, the operation is cumbersome, and the technical requirements for experimental operators are high, which is not conducive to the large-scale promotion of this project detection

Method used

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  • Kit for detecting concentration of oxidized low-density lipoprotein in sample and preparation method thereof
  • Kit for detecting concentration of oxidized low-density lipoprotein in sample and preparation method thereof
  • Kit for detecting concentration of oxidized low-density lipoprotein in sample and preparation method thereof

Examples

Experimental program
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Embodiment 1

[0034] Embodiment 1: preparation of biotin-labeled oxidized low-density lipoprotein clone antibody

[0035] (1) Activation of biotin

[0036] Biotin must be activated before coupling with the oxidized low-density lipoprotein clone antibody. The specific activation steps are as follows: Weigh 25 g of biotin and dissolve it in 30 ml of dimethylformamide solution, and then add N-succinimide ester 1.5 g and 2.0 g of biscyclohexyl carbodiimide, and stirred under airtight magnetic force at room temperature for 20-24 hours to precipitate a precipitate; the white precipitate was removed by suction filtration, and washed several times with dimethylformamide dropwise, and the filtrate was stored at 0°C Overnight, if the white precipitate is precipitated again, it should be treated by suction filtration again; the filtrate from which the white precipitate has been removed is heated to about 100°C, and the dimethylformamide in the solvent is removed by suction filtration; the obtained sol...

Embodiment 2

[0048] Example 2: Preparation of biotin-labeled oxidized low-density lipoprotein clone antibody

[0049] The preparation steps in this embodiment are basically the same as in Example 1, except that:

[0050] In step 5 of step (3), add 50% redistilled glycerol to the biotin-bound oxidized low-density lipoprotein clone antibody solution, and store at -15°C.

Embodiment 3

[0051] Example 3: Preparation of biotin-labeled oxidized low-density lipoprotein clone antibody

[0052] The preparation steps in this embodiment are basically the same as in Example 1, except that:

[0053] (2) Purification of oxidized low-density lipoprotein clone antibody:

[0054] Dilute the oxidized low-density lipoprotein clone antibody to be labeled at a concentration of 5 mg / ml with 100 mM carbonate solution to a concentration of 2 mg / ml, and dialyze with the diluted carbonate buffer for 15 hours.

[0055] (3) Coupling of activated biotin and oxidized low-density lipoprotein cloned antibody

[0056] The specific preparation method is as follows:

[0057] 1. Dissolve activated biotin in pure dimethyl sulfoxide at a final concentration of 5 mg / ml;

[0058] 2. Dissolve the purified oxidized low-density lipoprotein clone antibody to be coupled in 0.05mol / L pH9.0 sodium bicarbonate solution at a concentration of 5 mg / ml;

[0059] 3. Mix the activated biotin with the oxi...

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Abstract

The invention belongs to the technical field of external immunodetection and relates to a kit for detecting a concentration of oxidized low-density lipoprotein in a sample and a preparation method thereof. The kit comprises an anti-oxidized low-density lipoprotein monoclonal antibody-coated platform carrier, oxidized low-density lipoprotein concentration gradient standard substances with different concentrations, an oxidized low-density lipoprotein quality control product, a biotin-labeled oxidized low-density lipoprotein clonal antibody, a horse radish peroxidase-labeled avidin, a sample buffer, a substrate A, a substrate B, a stopping solution and a condensed washing lotion. The used detection devices are simple and are common devices so that the kit has a low detection cost and is conducive to popularization. The kit can be operated easily and is free of professional operators. A unique standard substance diluent in the kit and a sample buffer formula greatly improve detection result reliability and accuracy.

Description

technical field [0001] The invention belongs to the technical field of in vitro immunoassay, and in particular relates to a kit for detecting the concentration of oxidized low-density lipoprotein (Ox-LDL) in a sample and a preparation method thereof. Background technique [0002] Lipoprotein is the main carrier of plasma cholesterol and triglyceride; low-density lipoprotein (LDL) is a low density (1.019 ~ 1.063g / cm 3 ) plasma lipoprotein, composed of phospholipids, apolipoprotein (ApO) B polypeptide chain, cholesterol, triglycerides, carbohydrates, etc., containing about 25% protein and 49% cholesterol and cholesterol ester; oxidized low-density lipoprotein (OxLDL ) is a lipoprotein formed by natural low-density lipoprotein (LDL) through oxidative modification. [0003] In recent years it has been found that OxLDL is closely related to atherosclerosis. Ox-LDL promotes the formation of foam cells, and the death of foam cells will lead to the deposition of lipids on the arte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/92G01N33/577G01N33/543G01N33/531
CPCG01N33/92G01N33/531G01N33/543G01N33/577
Inventor 郑乐民张立峰李晓燕马志军吴建榕马奕晶
Owner 北京协和洛克生物技术有限责任公司
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