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Myoglobin detecting reagent

A technology of myoglobin and detection reagents, which is applied in the field of immunoassay, can solve the problems of poor batch-to-batch variation and inability to quantify stability, and achieve the effects of good stability, simple and fast detection, and improved detection range

Inactive Publication Date: 2017-11-24
QINGDAO CHENDA BIOLOGICAL SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of the problems still existing in the myoglobin detection reagent or method in the prior art, the myoglobin detection reagent of the present invention is proposed. The myoglobin detection reagent forms a composite material with polystyrene particles and nano gold (colloidal gold), It overcomes the shortcomings of the colloidal gold method that cannot be quantified and has poor stability (big differences between batches), and effectively improves the detection sensitivity and anti-interference ability, especially suitable for the determination of myoglobin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0020] Preparation of nano-gold-polystyrene particles

[0021] Under the protection of nitrogen, disperse 1g of dry polystyrene (the average particle size of polystyrene is 100nm) in 500ml of water, drop into 800ml of 0.8mmo / L HAuCl 4 Aqueous solution, stirred and reacted for 5 to 7 hours, then added 80ml of 8mmol / L sodium citrate aqueous solution, and then dripped 400ml of 0.015mol / L NaBH 4 After the aqueous solution is dropped, continue to stir and react for 2 to 4 hours, stand still, separate the solid from the liquid, and freeze-dry the obtained solid to obtain nano-gold-polystyrene particles. The average particle diameter of the nano-gold-polystyrene particles is measured to be 125nm.

preparation example 2

[0023] Preparation of nano-gold-polystyrene particles coated with myoglobin antibody

[0024] 1) Disperse the nano-gold-polystyrene particles obtained in Preparation Example 1 in glycine-sodium hydroxide with a pH of 9, and the concentration of the nano-gold-polystyrene particles is 10-20 mg / mL;

[0025] 2) Add the myoglobin antibody to the solution in step 1), stir overnight at room temperature, and centrifuge the solid-liquid to obtain nano-gold-polystyrene particles coated with the myoglobin antibody, which are stored for future use.

Embodiment 1

[0027] A detection reagent for detecting myoglobin, wherein the detection reagent for myoglobin includes reagent diluent, nano gold-polystyrene particles coated with myoglobin antibody, and the reagent diluent includes 50mM pH7.2~7.8 Tris-HCl, 0.8 wt% BSA, 0.05 wt% NaN 3 , 8% by weight of glycerin and 90mM NaCl; the average particle size of the nano-gold-polystyrene particles is 125nm; the concentration of the nano-gold-polystyrene particles coated with myoglobin antibody is 0.9mg / mL.

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Abstract

The invention discloses a myoglobin detecting reagent. The myoglobin detection reagent comprises a reagent diluent and nanogold-polystyrene particles coated with a myoglobin antibody, wherein the reagent diluent is prepared from 10-100 mM Tris-HCl with pH being 7.2-7.8, 0.5wt%-2wt% of BSA, 0.05wt%-0.1wt% of NaN3, 2wt%-15wt% of glycerol and 50-300 mM NaCl; the average particle size of the nanogold-polystyrene particles is 120-150 nm; the concentration of the nanogold-polystyrene particles coated with the myoglobin antibody is 0.5-2 mg / mL. The myoglobin detecting reagent has the advantages of good stability, simple and fast detection, high sensitivity and high anti-interference capacity, and is suitable for industrial application and popularization.

Description

technical field [0001] The invention belongs to the field of immunoassay, and in particular relates to a detection reagent for detecting myoglobin. Background technique [0002] Myoglobin (Myoglobin) is a specific oxygen-binding protein of skeletal muscle and cardiac muscle. The molecular weight is 16700, and it is composed of a polypeptide chain containing 153 amino acid residues and a heme prosthetic group, and is in a compact spherical shape. [0003] Due to the small molecular weight of myoglobin, when muscle cells are damaged, myoglobin will be released into the circulatory system. It rises in 1-2 hours, reaches the peak in 12 hours, and gradually decreases after 24 hours. It is an early diagnosis of acute myocardial infarction and other diseases. important reference indicators. [0004] At present, the commonly used immunoassay methods for myoglobin mainly include latex immunoturbidimetry, chemiluminescence immunoassay, and colloidal gold immunochromatography. Among...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/532
CPCG01N33/68G01N33/532
Inventor 鲍文佳
Owner QINGDAO CHENDA BIOLOGICAL SCI & TECH
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