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Preparation method of immunomagnetic microparticle

A technology of magnetic particles and yolk antibodies, which is applied in the preparation methods of peptides, biochemical equipment and methods, DNA preparation, etc., can solve the problems of low separation specificity, low efficiency, complex affinity separation operation of biological macromolecules, etc., and achieve The effect of high immobilization capacity, convenient and fast operation, and shortening the time required for removal

Active Publication Date: 2014-04-16
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to provide an immunomagnetic particle based on egg yolk antibody, its preparation method and its application, which solves the technical problems of complex affinity separation operation, low separation specificity and low efficiency of biological macromolecules

Method used

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  • Preparation method of immunomagnetic microparticle

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Embodiment 1

[0042] Embodiment 1: It is the preparation process of the immunomagnetic particles based on the yolk antibody (IgY) of the anti-human antibody and the gold magnetic particles, and the specific process of the application of the immunomagnetic particles to the separation and removal of antibodies from biological samples such as serum / plasma:

[0043] 1] Preparation of immunomagnetic microparticles:

[0044]a. Immobilization of anti-human antibody yolk antibody on the surface of gold magnetic particles: Take 1ml of gold magnetic particle suspension and add it to a 2ml 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. The egg yolk antibody was formulated into a solution with a concentration of 1 mg / ml, 100-2...

Embodiment 2

[0050] Example 2 is the preparation process of immunomagnetic particles based on anti-human serum albumin IgY (yolk antibody) and gold magnetic particles, and the application of this immunomagnetic particle to the separation and removal of human serum albumin from biological samples such as serum / plasma process, as follows:

[0051] 1] Preparation of immunomagnetic microparticles

[0052] a. Immobilization of anti-human serum albumin yolk antibody on the surface of gold magnetic particles: take 4ml of gold magnetic particles into a 5ml capacity centrifuge tube, 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. Prepare the egg yolk antibody against human serum albumin into a solution with a concentration of 1mg / ml, take 1.5ml and add it to the gold m...

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Abstract

The invention relates to a preparation method and application of an immunomagnetic microparticle based on a yelk antibody (IgY). The yelk antibody (IgY) refers to an antibody separated from yelks of poultries such as chickens, ducks and gooses, and the magnetic microparticle refers to a magnetic microparticle with the particle diameter of 1 nm-200 mu m, superparamagnetism and high specific surface area. The application of the immunomagnetic microparticle comprises separation of large biological molecules such as protein, nucleic acid and the like in a biological sample, separation of cells and immunoprecipitation tests. In the preparation method, the yelk antibody (IgY) is immobilized on the surface of the magnetic microparticle so as to prepare the immunomagnetic microparticle based on the yelk antibody (IgY). The purpose of separating large biological molecules and cells from the biological sample is achieved by using affinity of large biological molecules and corresponding yelk antibodies thereof as well as superparamagnetism of immunomagnetic microparticles.

Description

technical field [0001] The invention relates to an immunomagnetic particle based on yolk antibody, its preparation method and its application. Background technique [0002] In the prior art, the most commonly used method for specific affinity separation of biomacromolecules is an immunoaffinity chromatography method using agarose or resin as a carrier and using mammalian-derived antibody IgG as an affinity ligand. However, this chromatographic method has two limiting factors: one is to separate biological macromolecules in the form of a chromatographic column, which requires a complicated pretreatment process, and requires a large reaction volume, which is not suitable for processing due to the large dilution of the sample. Small-volume samples; second, the specificity of mammalian-derived antibodies is not high, and will bind to rheumatoid factors, etc., resulting in poor separation specificity. Contents of the invention [0003] The purpose of the present invention is t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/22C07H1/06C12N15/10C12N5/00G01N33/53
Inventor 崔亚丽胡佳陈超
Owner XIAN GOLDMAG NANOBIOTECH
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