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Application of DNA immunosorbent in preparation of ds-DNA antibody detection reagents

An immunoadsorbent and antibody detection technology, which is applied in the application field of DNA immunosorbent, can solve the problem of not being able to specifically confirm the deposition and distribution of anti-ds-DNA antibodies

Active Publication Date: 2019-03-26
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, H&E staining and ordinary immunofluorescence cannot specifically confirm the deposition and distribution of anti-ds-DNA antibodies in kidney tissue

Method used

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  • Application of DNA immunosorbent in preparation of ds-DNA antibody detection reagents

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0129] Preparation of Example 1 DNA-MSNs

[0130] (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.

[0131] (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.

[0132] (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 white solid, w...

Embodiment 2

[0137] Example 2 DNA-SiO 2 preparation of

[0138] (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.

[0139] (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.

[0140] (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 ethanol and wa...

Embodiment 3

[0142] Example 3 Preparation of AS1411-MSNs

[0143] 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 a DNA immunosorbent in preparation of ds-DNA antibody detection reagents. The application of the DNA immunosorbent in the preparation of the anti-ds-DNA antibodydetection reagents has the advantages that the inventor discovers that the DNA immunosorbent can qualitatively analyze ds-DNA antibodies deposited in renal tissues and specifically confirm depositionand distribution of the ds-DNA antibodies in the renal tissues when being applied to immunofluorescence; the application of the DNA immunosorbent has a simple method and is easy to operate, and is suitable for preparation of the reagents for detecting and analyzing lupus nephritis and lupus erythematosus.

Description

technical field [0001] The invention belongs to the application of DNA immunosorbent, in particular to the application of DNA immunosorbent in the preparation of anti-ds-DNA antibody detection reagent. 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 pathog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533
CPCG01N33/533
Inventor 陈填烽贺利贞林智明刘婷张曦
Owner JINAN UNIVERSITY
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