Colloidal gold test paper for detecting trace quantity oxygenize low density lipoprotein by indirect competition method

A low-density lipoprotein and colloidal gold test paper technology, which is applied in the detection field of oxidized low-density lipoprotein, can solve problems such as individual differences, and achieve the effects of low cost, convenient portability, and high detection accuracy

Inactive Publication Date: 2008-05-21
李红玉 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are certain limitations in the naked eye's discrimination of color shades, so there is a limit to the sensitivity of the semi-quantitative function of this detection method, and individual differences may occur depending on the individual's different discrimination of color shades

Method used

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  • Colloidal gold test paper for detecting trace quantity oxygenize low density lipoprotein by indirect competition method
  • Colloidal gold test paper for detecting trace quantity oxygenize low density lipoprotein by indirect competition method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Preparation of Monoclonal Antibody to Oxidized Low Density Lipoprotein

[0027] Mice were immunized with oxLDL (purchased from Beijing Xiehe Sanyou Technology Development Company). Splenocytes from immunized mice were hybridized with mouse myeloma cells to prepare hybridoma cells. The hybridoma cells with high positivity were selected for cloning to prepare monoclonal hybridoma cells. Continue to culture for 20 generations, screen and finally establish a hybridoma cell line that can stably secrete monoclonal antibody.

[0028] Ascites is prepared by inoculating hybridoma cells into the peritoneal cavity of mice, and then monoclonal antibodies are prepared by extracting antibodies from the ascites.

Embodiment 2

[0030] Colloidal gold preparation and method for labeling monoclonal antibody

[0031] Colloidal gold particles were prepared by trisodium citrate reduction method. Take 0.01% HAuCl 4 Aqueous solution 100ml, heat to boil. Quickly add 2ml of 1% trisodium citrate in water as needed, continue to boil for about 5min, wine red appears. Leave it to cool, and thus make colloidal gold particles with a diameter of about 20 nm.

[0032] The colloidal gold optimal binding protein amount was determined by coagulation colorimetry. With 0.1Mol / L K 2 CO 3 Adjust the pH of the colloidal gold to 9.0, and divide into ten tubes, 1ml in each tube. After gradiently diluting the protein concentration, add it to the above ten tubes respectively, and add 1ml to each tube. After 5 minutes, add 1ml of 10% NaCl solution to each of the above tubes, mix well and let stand for 2 hours to measure the OD value. According to the OD value, the amount of antibody needed to stabilize 1ml of colloidal gol...

Embodiment 3

[0036] Preparation and assembly of colloidal gold test strips for detecting oxidized low-density lipoprotein

[0037] Sample absorbent pad: Cut the glass fiber into a size of 1.9cm×30cm for use.

[0038] Gold pad: Soak glass fibers cut into 0.5mm×30cm in a certain concentration of gold-labeled antibody solution for 2 hours, dry at 37°C for 1 hour, seal, and store at 4°C for later use.

[0039] Chromatographic membrane: use nitrocellulose membrane or cellulose acetate membrane, cut into 2.5cm×30cm size. A certain concentration of oxidized low-density lipoprotein is coated at 1 cm from the lower end of the membrane as a detection band with a bandwidth of 2 mm, and 1 mg / ml animal anti-mouse (mostly goat anti-mouse) IgG is coated at an upward interval of 0.5 cm as a quality control band. After natural drying in the shade, it was blocked with PBS containing 1% BSA, 0.01mol / L, pH 7.4 for 2 hours, dried in the shade at room temperature, and sealed for later use.

[0040] Absorbent ...

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Abstract

The present invention provides an indirect competition method for golden test paper for detecting micro-oxidation low density lipoprotein colloid. The competition method is applied to the test paper. A certain amount of golden dot antibody is covered on the golden dot antibody pad. The oxidation low density lipoprotein is covered on a detecting region. Through the observation of the color of the detecting region, whether the oxidation low density lipoprotein in the serum or the plasma of a human body being higher than a normal level can be detected. The present invention is convenient and quick; the detection of the oxidation low density lipoprotein can be directly carried out; and the present invention provides reliable evidence to the early diagnose of atherosclerosis.

Description

1. Technical field: [0001] The invention relates to the detection of oxidized low-density lipoprotein, in particular to a colloidal gold test paper for detecting oxidized low-density lipoprotein. 2. Background technology: [0002] According to the latest statistics of the American Heart Association in 2001, cardiovascular disease has become the number one killer of human beings. Atherosclerosis is the pre-pathological basis of most cardiovascular and cerebrovascular diseases, so the research on the pathogenesis, diagnosis and prevention methods of atherosclerosis has become a hot spot in the medical field. [0003] Lipoproteins are the major carriers of plasma cholesterol and triglycerides. Low-density lipoprotein (LDL) is composed of phospholipids, apolipoprotein (Apo) B polypeptide chain, cholesterol, triglycerides, carbohydrates, etc. Oxidized low-density lipoprotein (oxLDL) is formed by oxidative modification of LDL. In recent years, it has been found that oxLDL is cl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/558G01N33/577
Inventor 李红玉陈瑜
Owner 李红玉
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