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Magnetic molecularly imprinted nano-particle as well as preparation method and application thereof

A magnetic molecular imprinting and nanoparticle technology, applied in chemical instruments and methods, other chemical processes, etc., can solve the problems of poor biocompatibility and poor selectivity, and achieve uniform particle size, simple preparation method and few steps. Effect

Active Publication Date: 2015-02-18
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In summary, due to poor biocompatibility or insufficient selectivity, the existing materials cannot be directly used to enter living cells to capture and inhibit the activity of target protein molecules in cells.

Method used

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  • Magnetic molecularly imprinted nano-particle as well as preparation method and application thereof
  • Magnetic molecularly imprinted nano-particle as well as preparation method and application thereof
  • Magnetic molecularly imprinted nano-particle as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1. Preparation of Western Blot Magnetic Fluorescent Nanoparticles

[0042] The process of preparing Western imprinted magnetic fluorescent nanoparticles in the present invention is as follows: figure 1 As shown, it specifically includes the following steps:

[0043] 1) Synthesis of Fe with suitable size 3 o 4 SiO 2 Nanoparticles (wherein, the diameter of the magnetic core is 5-10nm, and the thickness of the silicon dioxide layer is 10-20nm), and the surface of the silicon dioxide has amino groups, such as figure 2 (b) shown.

[0044] 90 mg of dry hydrophilic amino-modified Fe 3 o 4 Magnetic nanoparticles (such as figure 2 (a)) was fully ultrasonically dispersed in 20 mL of deionized water, and was added dropwise to a mixed solution containing 462 mL of cyclohexane, 120 g of Triton X-100 and 96 mL of n-hexanol under vigorous stirring.

[0045] Add 0.45mL tetraethoxysilane (TEOS) dropwise, react for 30min, then add 1.8mL concentrated ammonia water dropwi...

Embodiment 2

[0056] Example 2: Selective inhibition of deoxyribonuclease in solution and direct detection of its activity with DNA fluorescent probes

[0057] Without SiO 2 Due to the positive charge of the amino groups on the surface of the encapsulated magnetic nanoparticles, and the weak positive charge on the surface of the unmodified polyethylene glycol magnetic molecularly imprinted polymer, both of them have non-specific adsorption to DNA fluorescent probes, which will interfere with DNA fluorescence. In situ detection of probes. After silicon-coating and polyethylene glycol modification, the magnetic molecularly imprinted polymer has good dispersibility and stability in aqueous solution, and it can be added to the system to be tested to specifically inhibit the target enzyme. At this time, it can be directly added DNA fluorescent probe was used to measure its activity.

[0058] In addition, when there are other enzymes in the system (example III (Exo III) as an example), directly...

Embodiment 3

[0073] Example 3. Western Blot Magnetic Fluorescent Nanoparticles Rapidly Introduced into Cells and Imaging under the Action of a Magnetic Field

[0074] The prepared magnetic molecularly imprinted polymer has good stability and biocompatibility, and can be quickly introduced into Hela cells through endocytosis under the action of a magnetic field and inhibit the activity of DNase I therein. Afterwards, streptolysin O (SLO) protein was used to form micropores on the cell membrane, and DNA fluorescent probes were introduced into the cells to perform fluorescence imaging of DNase I activity to prove the in situ inhibitory effect of molecularly imprinted nanoparticles on DNase I. The detection and comparison of the activity of Exo III shows that its activity is not affected by the DNase I molecularly imprinted polymer. The DNase I probe uses the same sequence as above, the Exo III probe is limited by the detection channel of the fluorescence microscope, the FAM fluorescent labeli...

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Abstract

The invention provides a magnetic molecularly imprinted nano-particle as well as a preparation method and an application thereof. The nano-particle can perform specific recognition, trapping, separation and activity inhibition on target protein in a solution in vitro, more importantly, the nano-particle can enter a living cell rapidly under the action of a magnetic field and perform in-situ combination and activity inhibition on the target protein in the living cell, distribution of the nano-particles in the cell can be traced through a fluorescence microscope after fluorescence labeling, and quantitation can be performed through detection of fluorescence intensity.

Description

technical field [0001] The invention belongs to the technical field of nanomaterial preparation, and relates to a magnetic-core double-shell type protein molecularly imprinted polymer nanoparticle and a preparation method thereof, and its application to capture and activity inhibition of target proteins in solutions, especially to target proteins in living cells Protein capture and activity inhibition. Background technique [0002] Molecularly imprinted polymers refer to polymers obtained by assembling functional monomers with template molecules and then removing template molecules. They have the advantages of low price, good stability, and specific recognition. Compared with natural antibodies and receptor molecules, molecularly imprinted polymers have many advantages: simple preparation, low cost, good stability, reusable, applicable to a variety of targets, easy to modify, etc. [0003] Based on the principle of interaction between molecularly imprinted polymers and temp...

Claims

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Application Information

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IPC IPC(8): C08F292/00C08F220/54C08F220/06C08F220/60C08F220/56C08F222/38C08F222/20C08F2/48C08F2/38B01J20/26B01J20/30
Inventor 赵美萍刘艺斌翟筠秋董建桐
Owner PEKING UNIV
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