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Method for removing high abundance protein using gold magnetic particle

A gold magnetic particle and high-abundance technology, which is applied in the preparation of test samples, material inspection products, biological testing, etc., can solve the problems of few types of removal and low removal efficiency, and achieve increased specificity, high removal efficiency, The effect of high immobilization capacity

Active Publication Date: 2011-09-14
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is to provide a method for removing high-abundance proteins such as albumin / antibodies in biological samples such as serum / plasma with gold magnetic particles, which solves the problem that the existing methods for removing high-abundance proteins have few types and low removal efficiency Shortcomings

Method used

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  • Method for removing high abundance protein using gold magnetic particle
  • Method for removing high abundance protein using gold magnetic particle

Examples

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Embodiment 1

[0025] Embodiment 1 is the immobilization process of protein A on the surface of gold magnetic particles and the specific process of removing antibodies in biological samples such as serum / plasma with gold magnetic particles:

[0026]1] Immobilization of protein A on the surface of gold magnetic particles and sealing of gold magnetic particles: Take 1ml of gold magnetic particle suspension and add it to a 2ml capacity centrifuge tube, shake gently to resuspend. Place on a magnetic separator, magnetically separate, and discard the supernatant. Add 1ml of coupling buffer (0.2M Tris-HCl buffer at pH 8.0), shake gently to resuspend. Place on a magnetic separator, magnetically separate, and discard the supernatant. Repeat the operation once. Prepare protein A into a solution with a concentration of 1 mg / ml, add 100-200 μl to gold magnetic particles, and react in a shaker at 37°C and 180 r / min for 30 minutes. After the reaction is completed, magnetic separation is performed, and ...

Embodiment 2

[0031] Embodiment 2 is the specific process of immobilizing human serum albumin antibody on the surface of gold magnetic particles and using it to remove albumin in biological samples such as serum / plasma:

[0032] 1] Immobilization of human serum albumin antibody on the surface of gold magnetic particles and sealing of gold magnetic particles: take 4ml of gold magnetic particles into a centrifuge tube with a capacity of 5ml, shake gently to resuspend. Place on a magnetic separator, magnetically separate, and discard the supernatant. Add 4ml of coupling buffer and shake gently to resuspend. Place on a magnetic separator, magnetically separate, and discard the supernatant. Repeat the operation once. The human serum albumin antibody was formulated into a solution with a concentration of 1mg / ml, 1.5ml was added to the gold magnetic particles, and reacted in a shaker at 37°C and 180r / min for 30min. After the reaction is completed, magnetic separation is performed, and the super...

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Abstract

Said gold magnetic fine grains are made by mean of covering gold or silver metal on surface of the magnetic nanometer particle core, or by means of assembling the nanometer gold or silver metal particles on the surface of magnetic nanometer particle core. With the affinity chromatography method, the affinity ligand of the high abundant protein is fixed on the gold magnetic fine grains; the affinity interaction between the albumin and / or antibody and its affinity ligand, and the superparamagnetism of the gold magnetic fine grain are used for separating the albumin and / or antibody from the blood serum.

Description

technical field [0001] The invention relates to a method for removing high-abundance proteins in biological samples such as serum / plasma. Background technique [0002] Biological samples such as human serum / plasma contain a variety of low-abundance proteins secreted by tissue cells and shed into the blood, which may be key proteins in signal transduction and regulation. For example, during necrosis, apoptosis or hemolysis, some intracellular proteins are released into the blood. Carrying out serum proteomics research on diseases and looking for low-abundance proteins will help clarify the pathogenesis, and may discover some marker proteins associated with specific states of the disease, thereby providing new methods for disease diagnosis. The research results of serum proteomics can be applied to the clinical diagnosis of diseases, disease course monitoring, curative effect monitoring and drug treatment. However, the presence of some high-abundance proteins, especially alb...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/34G01N33/50
Inventor 陈超崔亚丽唐一通李铮
Owner XIAN GOLDMAG NANOBIOTECH
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