Method for preparing protein magnetic blotting nanospheres

A magnetic nanosphere and magnetic nanoparticle technology, applied in chemical instruments and methods, other chemical processes, etc., can solve the problems of unfavorable wide application, low adsorption capacity, large particle size of silicon spheres, etc., and achieve high coupling ligand capacity. , the effect of large specific surface area and small particle size

Inactive Publication Date: 2009-01-21
NANKAI UNIV
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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.

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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

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preparation example Construction

[0032] The preparation method of the protein magnetically imprinted nanosphere of the present invention is a method for preparing the protein magnetically imprinted nanosphere using one of hemoglobin, lysozyme, and serum albumin as a template molecule, including the following stages:

[0033] (1) Synthesis of magnetic nanoparticles;

[0034] It adopts the method of co-precipitation to synthesize magnetic nanoparticles, and the basic reaction equation of the co-precipitation method is: Fe 2+ +2Fe 3+ +8OH - = Fe 3 o 4 +4H 2 O, and the magnetic Fe is obtained by the following steps 3 o 4 Nanoparticles:

[0035] (1) FeCl 2 4H 2 O and FeCl 3 ·6H 2 O is placed in secondary water deoxidized by nitrogen gas, stirred and dissolved, and Fe 3+ / Fe 2+ =2:1 (molar ratio);

[0036] (2) Raise the ambient temperature to above 80°C, add ammonia solution drop by drop under nitrogen atmosphere, and make the OH - : Fe 3+ The molar ratio is greater than 15, and the mechanical stirr...

Embodiment 1

[0055] Example 1: Preparation of hemoglobin magnetic nanoimprinted polymer spheres

[0056] 1. Synthesis of superparamagnetic Fe by co-precipitation method 3 o 4 Preparation of nanoparticles

[0057] Basic reaction equation: Fe 2+ +2Fe 3+ +8OH - = Fe 3 o 4 +4H 2 o

[0058] (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);

[0059] (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

[0060] colored magnetic fluid;

[0061] (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

[0078] Example 2: Preparation of Lysozyme Magnetic Nanoimprinted Polymer Balls

[0079] The template protein was added as lysozyme, and the others were the same as in Example 1 to obtain lysozyme magnetic nanoimprinted polymer balls.

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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 and nucleic 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 Applications(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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