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Solid-phase synthesis method for nano silicon dioxide particles based on histidine tag

A nano-silica and histidine-labeled technology, which is applied in the direction of silica, silicon oxide, nanotechnology for materials and surface science, etc., can solve the problems of difficult separation and purification, difficult sample recovery, etc., and achieve convenient purification , The effect of simplifying the preparation process and improving the utilization rate

Active Publication Date: 2015-09-02
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to invent a solid-phase synthesis method of nano-silica particles based on histidine-tagged proteins in view of the shortcomings of the existing traditional preparation methods, which are difficult to separate and purify, and difficult to recover samples after use.

Method used

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  • Solid-phase synthesis method for nano silicon dioxide particles based on histidine tag
  • Solid-phase synthesis method for nano silicon dioxide particles based on histidine tag
  • Solid-phase synthesis method for nano silicon dioxide particles based on histidine tag

Examples

Experimental program
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Embodiment 1

[0024] Example 1: Preparation of red fluorescent protein (RFP) nano-silica particles based on histidine tag

[0025] First, resin particles containing nickel ions on the surface and red fluorescent protein based on histidine tag (can be replaced by green fluorescent protein GFP, near-infrared fluorescent protein IFP, yellow fluorescent protein YFP, cyan fluorescent protein CFP and other histidine-labeled protein, etc.) according to 0.05-0.25mg protein / mg resin particles into the phosphate buffer solution of pH = 7.0-8.0, stirred to make it fully mixed and then added to the gravity settling column with filter membrane. Add (3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) in a mass ratio of 1:2-5:2 for deionization In water, shake briefly and add to the gravity column, shake for 30-60min to complete the activation of the red fluorescent protein surface. Add 5-10 μL of 3-aminopropyltriethoxysilane, shake fully to dissolve it, react f...

Embodiment 2

[0027] Example 2: Further functionalization of red fluorescent protein (RFP) nano-silica particles based on histidine tag

[0028] The surface of the nano-silica particles re-adsorbed by the resin particles in Example 1 can be further functionalized according to the requirements: add 5-10 μL of 3-aminopropyltriethoxysilane, shake it fully to dissolve it, and react 12- After 16 hours, rinse the gravity column with deionized water to remove incompletely reacted cross-linking agent and reaction by-products. After that, 0.01-0.02mM FITC was added, and after fully shaking, it was placed in a shaker and shaken at a constant speed for 8-12h. Rinse the column again with deionized water to remove incompletely reacted FITC. After a simple elution process and removal of imidazole in the solution, the FITC-labeled nano-silica particles were obtained.

Embodiment 3

[0029] Example 3: Preparation of catalase nano-silica particles based on histidine tag

[0030]First, add the resin particles containing nickel ions on the surface and the catalase based on histidine tag into the phosphate buffer solution with pH=7.0-8.0 according to 0.05-0.5 mg enzyme / mg resin particles, stir to make them fully mixed and then add to Gravity settling column with filter membrane. According to the preparation method in Example 1, it is possible to obtain silica-coated catalase nanoparticles with an average particle size of 20nm, uniform particle size, and good dispersion, and the activity of catalase encapsulated by silica It can still maintain more than 90% of the original activity, and has more excellent stability. At the same time, the histidine tag on the surface of the nanoparticles enables it to be recycled many times.

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Abstract

The invention discloses a solid-phase synthesis method for nano silicon dioxide particles based on a histidine tag. The method comprises the following steps: firstly complexing with nickel ions on resin particle surfaces by virtue of the histidine tag of histidine tag protein; adsorbing the histidine tag protein on the resin particle surfaces; and growing the nano silicon dioxide particles employing immobilized protein as nucleus. The method has significant characteristics and advantages that nano particles are always kept in non-covalent link through the histidine tag and resin in the overall synthesis process of the nano silicon dioxide particles and the subsequent surface functionalization processes; the purification process after the reaction in each step is greatly simplified into a simple leaching process; the non-covalent link is reversible; after synthesis and functionalization are finished, the nano particles can be eluted from the resin particle surfaces though an eluant; and meanwhile, the eluted nano particles with the histidine tag can also be recovered through adsorption of the resin particles after being applied.

Description

technical field [0001] The invention relates to a solid phase synthesis method of nano silicon dioxide particles, in particular to a solid phase synthesis method of nano silicon dioxide particles based on histidine tags. Background technique [0002] Nanoparticles refer to microscopic particles of the nanometer scale, which have surface effects, small size effects, and quantum effects, and exhibit many new and different properties, that is, their optical, mechanical, thermal, and other chemical properties and bulk solids. There will be a noticeable difference. Nanoparticles are widely used in fields including energy, materials, biomedicine, and the environment due to their excellent potentials such as small particle size, large specific surface area, and good dispersion. [0003] As a special nanoparticle, nano-silica particles have many advantages while having the universal characteristics of nanoparticles: 1) As a non-toxic, odorless, and pollution-free non-metallic mater...

Claims

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Application Information

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IPC IPC(8): C01B33/18B82Y30/00
Inventor 曹傲能崔阳冬韩凯曹玉叶王海芳
Owner SHANGHAI UNIV
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