Time-resolved immunochromatographic kit for quantitatively detecting Ox-LDL (Oxidized Low-Density Lipoprotein) and application

A low-density lipoprotein, time-resolved technology, applied in the analysis of materials, biological testing, material stimulation analysis, etc., can solve the problem of large analytical instruments, high cost of reagents, poor sensitivity of Ox-LDL, not suitable for single-serve instant operation, etc. problem, to achieve the effect of wide detection range, overcome instability and save time

Pending Publication Date: 2020-02-04
北京协和洛克生物技术有限责任公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Several methods for measuring oxidized low-density lipoprotein include: 1) Traditional conjugated diene method, thiobarbituric acid chemical method, relative electrophoretic mobility method, ELISA method, etc. Long, complicated operation and not suitable for single-person instant operation, the sample is easily oxidized and leads to inaccurate shortcomings; 2) latex-enhanced turbidimetry is a monoclonal antibody cross-linked on the surface of polymer latex microspheres, while When the antigen is combined with the microspheres cross-linked with antibodies, they can quickly gather together in a short time and change the absorbance of the reaction solution to determine the content of Ox-LDL in the sample. However, this method has sensitivity to detect Ox-LDL Poor shortcomings, difficult to meet clinical testing needs
3) The chemiluminescence method has high sensitivity, but the operation is cumbersome, requiring special luminescent substrate-antigen / antibody conjugates, large analytical instruments, and high reagent costs, which limit its large-scale development and use.

Method used

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  • Time-resolved immunochromatographic kit for quantitatively detecting Ox-LDL (Oxidized Low-Density Lipoprotein) and application
  • Time-resolved immunochromatographic kit for quantitatively detecting Ox-LDL (Oxidized Low-Density Lipoprotein) and application
  • Time-resolved immunochromatographic kit for quantitatively detecting Ox-LDL (Oxidized Low-Density Lipoprotein) and application

Examples

Experimental program
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Effect test

Embodiment 1

[0078] This example is used to illustrate the preparation of the marker pad in the kit of the present invention.

[0079] (1) Selection of Eu fluorescent microspheres

[0080] Binding on the binding pad: antibody-labeled time-resolved Eu fluorescent microspheres are prepared by spraying gold buffer solution and diluting the labeled microspheres by 15 times, then spraying the microspheres on the binding pad according to the settings in Table 1, wherein spraying gold buffer solution Contains: 10mMTris-HCl+5% (w / v) sucrose+3% (w / v) trehalose+1% (w / v) BSA+0.1% (v / v) Tween 20+0.1% (v / v) Proclin 300. The time-resolved Eu fluorescent microspheres are polystyrene-carboxy PS(-COOH) microspheres with a particle size of 210nm, including no arms, surface-modified 6-atom straight-chain arms, surface-modified 200-atom straight-chain arms, and surface-modified 1000-atom There are four types of linear arms, preferably surface-modified 6-atom linear arm Eu fluorescent microspheres.

[0081...

Embodiment 2

[0097] This example is used to illustrate the preparation of nitrocellulose membrane (NC membrane) in the kit of the present invention.

[0098] (1) Nitrocellulose membrane scribing process There is an anti-APOB monoclonal antibody detection line T line and a goat anti-mouse polyclonal antibody quality control line C line on the nitrocellulose membrane, wherein the T line is coated with a coating solution ( In the present embodiment, it is also called the film-drawing diluent) to prepare the anti-APOB monoclonal antibody with a concentration of 0.8mg / ml, and the C line is to prepare the concentration of 0.5 with the coating solution (also known as the film-drawing diluent in this embodiment). mg / ml goat anti-mouse polyclonal antibody, and these two concentrations of antibodies were sucked into the pipeline of the film-drawing instrument, and the C line was drawn on the upper edge of the nitrocellulose membrane (NC membrane) by the film-drawing instrument, and the T line was dra...

Embodiment 3

[0104] This example is used to illustrate the preparation of the sample diluent of the kit of the present invention.

[0105] The sample diluent is prepared as Tris-HCl buffer solution of 0.05M pH 8.0, Proclin300 of 0.5% (v / v), 0.2% BSA, 0.2% (v / v) Tween-20, 0.5% (w / v) brij-35, 0.1 (w / v) Vitamin C.

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Abstract

The invention provides a time-resolved immunochromatographic kit for quantitatively detecting an Ox-LDL (Oxidized Low-Density Lipoprotein) and application. The kit comprises sample diluents, a test card and an ID card on which human Ox-LDL standard curve information is copied, wherein the test card consists of a PVC board, a sample pad, a marking pad, a conjugate pad, a nitrocellulose membrane andabsorbent paper; and Eu fluorescent microspheres for marking anti-Ox-LDL antibody are sprayed on the marking pad. The kit only needs to detect a 5ul blood sample at the normal temperature, and relative to conventional detection, has the characteristics of small blood sampling volume, rapidness, sensitivity, high accuracy, high specificity and high stability.

Description

technical field [0001] The invention belongs to the field of biomedical detection, and in particular relates to a rapid detection kit for quantitatively measuring the concentration of oxidized low-density lipoprotein (Ox-LDL) in human plasma by time-resolved immunochromatography. Background technique [0002] The structure of low-density lipoprotein (LDL) is an approximate spherical emulsion particle structure model. The spherical interior is neutral, consisting of cholesterol esters and a small amount of triglycerides; the outer layer is a hydrophilic and lipophilic monolayer, including phospholipids, unesterified cholesterol, and a molecule of apolipoprotein apoB-100. Phospholipids, cholesterol and proteins in LDL are very vulnerable to the attack of oxygen free radicals and hydroxyl free radicals and are oxidized into oxidized low-density lipoprotein (Ox-LDL). In vivo LDL can be oxidized in the following two ways: (1) arterial endothelial cells can be oxidized and modifi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/533G01N33/543G01N33/577G01N33/68G01N21/64
CPCG01N21/6408G01N33/5302G01N33/533G01N33/543G01N33/577G01N33/68
Inventor 吴建榕刘永黄龙耀丁琪王峥辉
Owner 北京协和洛克生物技术有限责任公司
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