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Composite material capable of enriching and immobilizing protein as well as preparation method and application thereof

A composite material and protein technology, which is applied in the field of preparation and application of nanomaterials, can solve the problems of weak activity of oxidase protein, large size, enrichment and enrichment ability of various proteins, etc. The method is simple, the ionic characteristics are improved, Efficient enrichment and fixed effects

Active Publication Date: 2021-03-30
XIAMEN HUAXIA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the existing Fe 3 o 4 Magnetic nano-functionalized modification / composite materials cannot enrich various proteins or have poor enrichment ability; the technical problems of weak activity of oxidase protein in catalytic reaction and large amount of enzyme body required, the present invention provides a kind of A composite material that enriches and immobilizes proteins. The composite material can achieve a variety of proteins in specific environments by modifying carboxylated ferroferric oxide magnetic nanospheres on bimetallic phosphate hydrate compounds with wrinkled sheet structures. High-efficiency enrichment and immobilization of oxidase proteins, and the reactivity of oxidase proteins can be significantly improved after immobilization, reducing the amount of enzymes used

Method used

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  • Composite material capable of enriching and immobilizing protein as well as preparation method and application thereof
  • Composite material capable of enriching and immobilizing protein as well as preparation method and application thereof
  • Composite material capable of enriching and immobilizing protein as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Synthesis of carboxylated tetrashydoxide three ferromagnetic nanospheres:

[0043] Accurately, the metal source hexahydrate 1.350g was added to a clean beaker and then 70 ml of solvent ethylene glycol was added, and the magnetic stirring (850 rpm) to hexahydrate was dissolved at room temperature. Then, a stabilizer citrate was added 0.24 g of citric acid and 3.85 g of reductant acetate, and stirring was continued for 20 minutes at room temperature to cover the fresh film at room temperature to obtain an emulsion. After the above emulsion was reacted in an oil bath at 140 ° C, it was transferred to a polytetrafluoroethylene reaction kettle, and after 16 h, a water was washed with ethanol and dried in vacuo to obtain a carboxylated tetraned iron magnetic nanosphere. .

Embodiment 2

[0045]Preparation of composite materials that can be enriched and fixed protein:

[0046] Accurately weigh 0.6435 g of itrate, zinc chloride 0.24 g is added to N, N-dimethylformamide, ultrasonic stirring dissolve, and an ammonia water 2 mL and obtained carboxylated Fe 3 O 4 Magnetic nanlur is 0.1g, and the ultrasonic treatment is mechanically stirred for 1 h, and the second precursor solution is obtained. The reaction system is obtained after obtaining the reaction system after 0.9 g of hydrogenmmonia and sodium citric acid, and the reaction system is added to the reaction system after stirring at room temperature for 10 min, and then the reaction system (solution) is placed again. After the polytetrafluoroethylene reaction kettle, after 24 hours at 180 ° C, the resulting material was magnetically separated and washed with water and ethanol, and then dried in vacuo drying 60 ° C for 2 h, resulting in the enrichment and fixation Composite of proteins.

Embodiment 3

[0048] Protein enrichment and fixation method:

[0049] 20 mg Fe 3 O 4 @Zn x Zr (PO 4 ) 2 YH 2 O Preparation of 10 mg / ml of dispersion, ultrasonic oscillating, forming a mixed solution of 10 mg / ml, and then taking 100 μL of the mixture, after the magnetic scattered aqueous solution, add a variety of different Protein solution (including human serum immunoglobulin Ig-g, chicken egg white protein OVA, transferrin TRF, bovine serum albumin BSA, hemoglobin HB, myoglobin MB, cytochrome C (Cyt-C), lactoferrin LF, human albumin HRP, lysozyme LYZ solution, shake at room temperature for 10 min, promotes protein adsorption in the surface of the composite to reach the enriched effect. 16 μg / ml of cytochrome C (Cyt-C) solution was added thereto, and the cytochrome C was immobilized at room temperature to promote the surface of the composite material to form a solution of a fixed protein.

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Abstract

The invention discloses a composite material capable of enriching and immobilizing protein as well as a preparation method and application thereof; the composite material comprises carboxylated ferroferric oxide magnetic nanospheres, which are modified on the surface of bimetallic phosphate hydrate with a sheet structure with wrinkles. The invention further discloses a preparation method of the composite material capable of enriching and immobilizing the protein. The preparation method comprises the following steps: firstly, obtaining carboxylated ferroferric oxide magnetic nanospheres; and then mixing the ferroferric oxide magnetic nanospheres obtained in the step S1 with the ZnxZr(PO4)2.yH2O to obtain the composite material. The composite material provided by the invention can realize efficient enrichment and immobilization of multiple proteins, and after immobilization of oxidase proteins, the reaction activity of the oxidase proteins can be significantly improved, and the use amount of enzymes is reduced.

Description

Technical field [0001] The present invention relates to the preparation and application of nanomaterials, in particular a composite material that enriches and fixes protein and its preparation methods and applications. Background technique [0002] With a variety of biological samples, the improvement of various instruments development and testing techniques have not met the treatment of complex samples and the selective separation analysis of multiple abundance proteins. Therefore, an effective pre-treatment method is taken and is essential for pre-separation enrichment of protein effective pre-treatment materials in conjunction with the appropriate sample. At present, the current nanomaterial adsorbent study is the weight of the research in the field of sample pre-treatment, and the nanomaterials have volume effects, surface effects, dielectric limits, and quantum dimensional effects, and therefore also imparted magnetic, thermal sensitivity, chemical activity, A variety of dif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22B01J20/28B01J20/30C12N11/14C12N9/02
CPCB01J20/22B01J20/06B01J20/28009B01J20/28021C12N11/14C12N9/0004
Inventor 姚秋虹郭志勇张晨陈曦
Owner XIAMEN HUAXIA UNIV
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