Method for preparing protein magnetic blotting nanospheres

A magnetic nanosphere and magnetic nanoparticle technology, applied in chemical instruments and methods, and other chemical processes, can solve the problems of low adsorption capacity, low specific surface area, and easy embedding in them, so as to avoid centrifugation and have wide application prospects , The effect of simplifying the centrifugal separation process

Inactive Publication Date: 2010-12-08
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the large particle size and relatively low specific surface area of ​​silicon spheres, some recognition sites are easily embedded in them, resulting in low adsorption capacity, and the use of silicon spheres as the separation medium involves cumbersome operations such as centrifugation, which is not conducive to wide application.

Method used

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  • Method for preparing protein magnetic blotting nanospheres
  • Method for preparing protein magnetic blotting nanospheres
  • Method for preparing protein magnetic blotting nanospheres

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1: Preparation of hemoglobin magnetic nanoimprinted polymer spheres

[0058] 1. Synthesis of superparamagnetic Fe by co-precipitation method 3 o 4 Preparation of nanoparticles

[0059] Basic reaction equation: Fe 2+ +2Fe 3+ +8OH - = Fe 3 o 4 +4H 2 o

[0060] (1) FeCl 2 4H 2 O (1.72g) and FeCl 3 ·6H 2 O (3.72g) was dissolved in 80mL of secondary water deoxidized by nitrogen, stirred and dissolved, wherein Fe 3+ / Fe 2+ =2:1 (molar ratio);

[0061] (2) Transfer the solution to a 250ml three-necked flask, heat it to 80°C under a nitrogen atmosphere, add 10ml of ammonia water (25%) dropwise, and mechanically stir (at a speed of 800rpm), and react for 30 minutes to obtain black

[0062] colored magnetic fluid;

[0063] (3) After cooling, the mixed solution is divided into two phases by magnetic separation with a magnet, and the supernatant is removed to obtain a black superparamagnetic Fe 3 o 4 Gel, washed twice with water to neutral, vacuum dried to...

Embodiment 2

[0080] Example 2: Preparation of Lysozyme Magnetic Nanoimprinted Polymer Balls

[0081] The template protein was added as lysozyme, and the others were the same as in Example 1 to obtain lysozyme magnetic nanoimprinted polymer balls.

Embodiment 3

[0082] Example 3: Preparation of Serum Albumin Magnetic Nanoimprinted Polymer Balls

[0083] Serum albumin lysozyme was added as the template protein, and the others were the same as in Example 1 to obtain serum albumin magnetic nanoimprinted polymer balls.

[0084] exist figure 1 In the effect figure of the ferroferric oxide magnetic nanoparticles in the shown embodiments 1, 2, and 3, the average particle diameter is 12nm; figure 2 In the effect diagram of the hemoglobin magnetic nanoimprinted polymer prepared in Example 1 shown, the average particle diameter is 100nm; image 3 Shown is the isothermal adsorption curve of the hemoglobin magnetic nanoimprinted polymer microspheres prepared in Example 1 on the template molecule hemoglobin.

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Abstract

The invention relates to a preparation method of a protein magnetic imprinting nano sphere, in particular to a preparation method of a protein magnetic imprinting nano sphere which takes one of hemoglobin, muramidase or serum protein as template molecules; the preparation method includes the following steps: synthesizing magnetic nano particles which are Fe3O4 nano particles covered with silicon dioxide on the surfaces; carrying out silanization to the amino-groups on the surface of the magnetic sphere; using glutaric dialdehyde to connect protein; using silylation agent to fix the space structure of template protein; alkaline eluting the protein on the surface of the magnetic sphere to form imprinting sites. The hemoglobin, muramidase or serum protein magnetic nano imprinting polymer sphere prepared by the invention combines the precise and specific identification property of molecular imprinting and the quick separation property of the magnetic nano sphere under the action of an external magnetic field, integrates the advantages of molecular imprinting and the magnetic nano sphere, avoids complex centrifugation operation, plays vital roles in the separation of cells, protein andnucleic acid, the detection of biomolecules, tumor diagnosis and medicine targeting therapy and has promising application prospect in the field of isolation analysis.

Description

technical field [0001] The invention relates to the preparation of a protein molecularly imprinted polymer. In particular, it involves a protein magnetic imprinting nanometer that has certain specific adsorption properties for template proteins, uses magnetism as a carrier, can be rapidly separated under the action of an external magnetic field, can avoid traditional complicated operations such as centrifugation, and simplifies the separation and recognition process. Ball preparation method. Background technique [0002] Molecular Imprinting Technique (MIT) is a technology for preparing polymer compounds (Molecular Imprinted Polymers, MIPs) with specific predetermined selectivity for specific target molecules (template molecules, also known as imprinted molecules) (M.D. Yan, O. Ramstrom, Ed., Molecularly Imprinted Materials Science and Technology, Marcel Dekker, New York, 2005). Molecularly imprinted polymers have holes whose shape matches the substrate molecules, and func...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/285B01J20/30
Inventor 陈朗星王新王莲艳何锡文张玉奎王彦芬
Owner NANKAI UNIV
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