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Application of silicon dioxide nanoparticles to DNA (deoxyribonucleic acid) immunoadsorbent

An immunoadsorbent and nanoparticle technology, which is applied in the direction of selective adsorption, inorganic chemistry, ion exchange, etc., can solve the problems of DNA immobilized capacity decrease, shedding, insufficient embedding, etc.

Active Publication Date: 2019-02-01
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there are two disadvantages of DNA immobilization by collodion embedding: one is that some DNA may fall off in the subsequent production process due to insufficient embedding, resulting in a decrease in the amount of DNA immobilized; DNA buried too deep and out of reach of antibodies

Method used

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  • Application of silicon dioxide nanoparticles to DNA (deoxyribonucleic acid) immunoadsorbent
  • Application of silicon dioxide nanoparticles to DNA (deoxyribonucleic acid) immunoadsorbent
  • Application of silicon dioxide nanoparticles to DNA (deoxyribonucleic acid) immunoadsorbent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0123] Preparation of Example 1DNA-MSNs Nanoparticles

[0124] (1) Ultrasonically disperse 4.0 g of cetyltrimethylammonium chloride (Aladdin Company) in 40 mL of water, add 106.8 mL of triethylamine (Aladdin Company) dropwise, stir for 1 hour, and add 3 mL of orthosilicon dropwise Acetate ethyl ester (Aladdin Company), stirred and refluxed at 90°C for 2h. After the reaction was completed, the samples were collected and centrifuged at 12000 rpm to obtain mesoporous silica nanoparticles.

[0125] (2) The mesoporous silica nanoparticles were washed three times with ethanol and ultrapure water respectively, dried in an oven at 60°C, ground into powder, and calcined in a muffle furnace at 550°C for 5 hours.

[0126] (3) Take 100 mg of mesoporous silica nanoparticles treated in step (2) and disperse them in 20 mL of toluene, add 2 mL of 3-aminopropyltriethoxysilane (Aladdin Company), stir for 30 min, heat up to 110 ° C and reflux Centrifuge at 12,000 rpm for 24 hours to obtain a w...

Embodiment 2D

[0129] Example 2 DNA-SiO 2 Preparation of nanoparticles

[0130] (1) Mix and stir 142.8mL ethanol, 20mL water and 3.14mL ammonia solution for 10min, then slowly add 6mL tetraethyl silicate (Aladdin Company), stir and react for 2h, centrifuge at 12000rpm, wash with water and ethanol three times respectively , freeze-dried to obtain solid silica nanoparticles.

[0131] (2) Disperse 100mg of solid silica nanoparticles in 20mL of toluene, add 2mL of 3-aminopropyltriethoxysilane (Aladdin Company), stir for 30min, heat up to 110°C and reflux for 24h, centrifuge at 12000rpm, wash with ethanol Wash with water three times respectively to obtain solid silica nanoparticles with amino groups on the surface.

[0132] (3) Disperse 100 mg of solid silica nanoparticles containing amino groups on the surface obtained in step (2) with ethanol, then add 400 mg of succinic anhydride (Aladdin) dispersed with ethanol, stir for 5 h at 37 ° C, and centrifuge at 12000 rpm , washed three times with ...

Embodiment 3A

[0134] Preparation of embodiment 3AS1411-MSNs nanoparticles

[0135] Take 50 mg of carboxyl-modified mesoporous silica nanoparticles (MSNs-COOH) obtained in step (4) of Example 1 and disperse them uniformly in 15 mL of water, and add 10 mg / mL N-hydroxysuccinimide (NHS, Aladdin Company) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC, Aladdin Company) each 2mL, stirred at room temperature for 2h, added Tris-HCl solution (0.05M , pH=8.0) dissolved AS1411 nucleic acid aptamer (purchased from Sigma, USA, nucleotide sequence shown in SEQ ID NO:1) solution (concentration is 2.5mg / mL, adding amount is 1mL), stirred at room temperature for 24h , centrifuged at 12000rpm, washed with water three times, freeze-dried, and ground to powder to obtain AS1411-MSNs nanoparticles.

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Abstract

The invention discloses application of silicon dioxide nanoparticles to DNA (deoxyribonucleic acid) immunoadsorbent. The application has the advantages that the application is based on the merits which are discovered by the inventor and include the good adsorption performance and the excellent safety performance of the DNA immunoadsorbent, and DNA is loaded in silicon dioxide nanoparticles to obtain the DNA immunoadsorbent; the DNA immunoadsorbent with the DNA loaded in the silicon dioxide nanoparticles is obtained on the basis of the application, the DNA is modified in the surfaces or pore passages of the nanoparticles MSNs and SiO2 the immobilization amount is high, falling can be prevented, and accordingly the safety of the immunoadsorbent can be improved; DNA molecules and anti-ds-DNA(double-stranded DNA) antibodies can be in sufficient contact with one another, the specific binding capacity can be realized to the greatest extent, and accordingly the prepared immunoadsorbent is good and stable adsorption performance.

Description

technical field [0001] The invention belongs to the field of blood purification, and in particular relates to the application of silicon dioxide nanoparticles in DNA immunoadsorbents. Background technique [0002] Systemic lupus erythematosus (SLE) is a chronic autoimmune disease with multi-system damage. The exact etiology is unknown, and it often causes irreversible damage to multiple organ systems. The central nervous system, etc., the pathological manifestations are the deposition of autoantibodies and immune complexes, which affect the life span and quality of life of patients. The prevalence of SLE varies from population to population, and it is more common in women in my country, especially women of childbearing age. A variety of antibodies represented by antinuclear antibody (ANA) and anti-double-stranded DNA antibody (anti-ds-DNA antibody) appeared in the patient's serum, among which anti-ds-DNA antibody is the main antibody involved in the pathogenesis of SLE, and...

Claims

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Application Information

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IPC IPC(8): B01J20/10B01J20/30B01D15/08
CPCB01D15/08B01J20/103
Inventor 陈填烽林智明
Owner JINAN UNIVERSITY
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