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Immune turbidimetry for detection of apoliprotein M

An apolipoprotein and turbidimetric technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problem of not having a single measurement method, and achieve the effects of reducing detection cost, low price and simple operation.

Inactive Publication Date: 2004-12-22
张晓膺
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] To sum up, the determination of the content of ApoM in plasma or serum has a wide range of clinical application prospects; at the same time, because ApoM is a newly discovered protein, its function needs to be further studied. In the study, the specificity is strong. , high sensitivity, good stability, simple and easy method for measuring ApoM in human plasma or serum is an indispensable tool, but there is still no applicable assay method so far

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1 (with pure ApoM as standard substance)

[0046] 1. Prepare the following reagents and instruments

[0047] ①, diluent: take 0.2g KH 2 PO 4 , 2.9g Na 2 HPO 4 12H 2 O, 8g NaCl, 0.5ml Tween20, 40g polyethylene glycol-6000, add deionized water to 1000ml, mix evenly and filter with a 5G glass core funnel for later use, and store it in a refrigerator at 2-6°C for half a year. The absorbance A340 of this diluent is less than 0.05, and this value will still be maintained after half a year, and it can be used continuously. If it is greater than 0.05, it can be used after suction filtration with a 5G glass core funnel;

[0048] ②. The precision photometer is ultraviolet-visible spectrophotometer 751 or the automatic analyzer is OLYMPUS large-scale automatic biochemical analyzer AU2700; the turbidimeter uses Beckman turbidimeter.

[0049] 2. Prepare anti-ApoM polyclonal antibody

[0050] The anti-ApoM polyclonal antibody is diluted 1:200 with diluent for detect...

Embodiment 2

[0055] Embodiment 2 (make standard substance with pure ApoM)

[0056] Items 1, 2, and 3 are operated according to Example 1, and then on the AU2700 type automatic analyzer, set the sample volume (i.e., the sample to be tested) to 40 μl, the reagent volume (i.e., the anti-ApoM polyclonal antibody) to 160 μl, and the reaction temperature The temperature is 37°C, the reaction time is 30 minutes, and the detection wavelength is 340nm. Use the prepared ApoM standard for calibration, and use the diluent as a blank to detect the ApoM content of the sample to be tested, and the instrument will automatically give the detection result.

Embodiment 3

[0057] Embodiment 3 (with pure ApoM as standard substance)

[0058] Except that the AU2700 automatic analyzer is replaced by a turbidimeter (such as a Beckman turbidimeter), and the light scattering intensity rate method is performed immediately after the sample to be tested is set to react with the anti-ApoM polyclonal antibody, the rest of the operations are carried out in the same way. Example 2.

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PUM

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Abstract

In the method, reaction between polyclonal antibody of anti apolipoprotein M and apolipoprotein M of sample to be tested forms compound, which causes change of cloudiness. The change is expressed by intensity of light transmission or intensity of light scattering. Content of apolipoprotein M of sample to be tested is looked out from curve between concentrations of standard apolipoprotein M and relevant intensity of light transmission or intensity of light. Titer of polyclonal antibody is at least 1:128 containing multi antigenic determinant. Sample to be tested is human blood plasma or serum in empty stomach diluted. Polyclonal antibody of anti apolipoprotein M is mixed with diluted sample. Features of thee method are: convenience, good repeatability, and high adaptability.

Description

technical field [0001] The invention relates to an immunoturbidimetric method for detecting apolipoprotein M (Apoliprotein M, referred to as ApoM), and more specifically, relates to a method for measuring ApoM content in human plasma or serum. Background technique [0002] Lipoprotein is a macromolecular complex that transports various lipids in blood and other body fluids, participates in various physiological and pathological processes in the body, and is especially closely related to lipid metabolism disorders in the body and the occurrence and development of cardiovascular diseases. There are different types of lipoproteins, which are currently classified according to their physical and chemical properties, the most important of which is based on their density during ultracentrifugation. According to this, it can be divided into chylomicrons (CM, density<1.006), very low-density lipoprotein (VLDL, density<1.006), intermediate density lipoprotein (IDL, 1.006<dens...

Claims

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Application Information

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IPC IPC(8): G01N33/536
Inventor 徐宁
Owner 张晓膺
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