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Magnetic nanoparticle biological probe, preparation method and application thereof

A technology of magnetic nanoparticles and biological probes, applied in the fields of biotechnology and nanomaterials, can solve the problems of high cost of detection experiments, inability to quantitatively detect HBV, and long time consumption.

Inactive Publication Date: 2015-09-16
SUZHOU WIN BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to overcome the "two pairs of half" reagents in the prior art can not well reflect the existence of hepatitis B virus, can not quantitatively detect HBV, and HBV-DNA quantitative PCR detection Due to the high cost and time-consuming defects of the experiment, the present invention provides a magnetic nanoparticle bioprobe for detecting HBV preS1 antigen, a magnetic immunochromatography test strip, a preparation method and a use method thereof, and the magnetic nanoparticle bioprobe With the characteristics of strong specificity, sensitive signal and good stability, its magnetic immunochromatographic test strip can realize the rapid qualitative and quantitative detection of HBV preS1 antigen in human peripheral serum

Method used

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  • Magnetic nanoparticle biological probe, preparation method and application thereof
  • Magnetic nanoparticle biological probe, preparation method and application thereof
  • Magnetic nanoparticle biological probe, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 magnetic nanoparticles

[0074] A method for preparing magnetic nanoparticles, comprising the steps of:

[0075] (1) Prepare magnetic particles by solvothermal method: Dissolve 0.85g FeCl3 6H2O and 0.4033g PAA in 40ml ethylene glycol respectively, then add 1.2g urea to dissolve, mix and add to a stainless steel reactor with tetrafluoroethylene liner In the process, tighten the reactor, heat and react at 220°C for 16 hours, and wash with ethanol and deionized water three times with the assistance of magnetic separation to obtain magnetic particles with an average particle size of 90nm;

[0076] (2) Silicon oxide-coated magnetic particles: Mix 90 mg of magnetic particles with 1.0 mol / L hydrochloric acid solution, sonicate for 30 minutes, wash 5 times with deionized water under the assistance of magnetic separation, and then mix the magnetic particles with alcohol water volume Mix 125 mL of alcohol-water mixture with a ratio of 70:30, add 200 μL of tetraethyl...

Embodiment 2

[0078] Example 2 magnetic nanoparticles

[0079] A method for preparing magnetic nanoparticles, comprising the steps of:

[0080] (1) Prepare magnetic particles by solvothermal method: Dissolve 0.81g FeCl3 6H2O and 2.66g sodium citrate in 30ml ethylene glycol respectively, mix them and add them to a stainless steel reaction kettle with a tetrafluoroethylene liner, tighten the reaction Kettle, heated and reacted at 200°C for 24 hours, washed with ethanol and deionized water three times with the assistance of magnetic separation, to obtain magnetic particles with an average particle size of 250 nm;

[0081] (2) Silicon oxide-coated magnetic particles: Mix 110 mg of magnetic particles with 1.0 mol / L hydrochloric acid solution, sonicate for 15 minutes, wash with deionized water for 7 times with the assistance of magnetic separation, and then mix the magnetic particles with alcohol water volume Mix 125 mL of alcohol-water mixture with a ratio of 90:10, add 100 μL of tetraethyl ...

Embodiment 3

[0083] Example 3 Magnetic Nanoparticle Bioprobes

[0084] A method for preparing a magnetic nanoparticle bioprobe, comprising the steps of:

[0085] Step 1) Add 15 mg of the magnetic nanoparticles prepared in Example 1 or 2 to 50 g of ethanol aqueous solution with a volume ratio of alcohol to water of 70:30, add 50 μL of ammonia water and 35 μL of aminopropyltriethoxysilane, and stir for 2 hours at room temperature , continue to stir for 3 hours in a water bath at 70°C, wash with ethanol to obtain surface-aminated magnetic nanoparticles; mix the surface-aminated magnetic nanoparticles with dimethyl sulfoxide (DMSO), add 5.5 mg of succinic anhydride to react for 24 hours , washed three times with DMSO under the assistance of magnetic separation, and then washed three times with deionized water to obtain surface carboxylated magnetic nanoparticles, the carboxyl content of these surface carboxylated magnetic nanoparticles is 0.9-1.5mmol / g;

[0086] Step 2) Wash 2 mg of surfac...

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Abstract

The invention discloses a magnetic nanoparticle biological probe, a preparation method and application thereof, in particular to a magnetic nanoparticle biological probe for detecting HBV preS1 antigen, a magnetic immunochromatography test paper strip, a preparation method and a use method thereof. According to the invention, magnetic nanoparticles with carboxyl modified surfaces are adopted as the carrier, the antibody of HBV preS1 is connected to the magnetic nanoparticles' surfaces through EDC and HBV to establish the preparation method of the magnetic nanoparticle biological probe for detecting the HBV preS1 antigen, and the biological probe is adopted as the marker and the antibody of the targeted marker is taken as the capture antibody to establish a sandwich type immunochromatography detection system. The magnetic nanoparticle biological probe has strong specificity, and has good signal sensitivity and stability. The magnetic immunochromatography test paper strip provided by the invetnion on the one hand can realize rapid qualitative detection of the HBV preS1 antigen through visual inspection, and on the other hand can realize quantitative detection of the HBV preS1 antigen through the quantitative magnetic signal of the captured probe.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and nanomaterials, and in particular relates to a magnetic nano particle biological probe for detecting HBV preS1 antigen, a magnetic immunochromatographic test strip, a preparation method and a use method thereof. Background technique [0002] Hepatitis B (HBV) is a high incidence in my country. The infection rate among the Chinese population is 60%, and it is increasing year by year. There are more than 130 million HBV surface antigen carriers, and more than 10% of them turn into chronic hepatitis, and some of them develop into liver cirrhosis. , leading to liver cancer. Therefore, it is particularly important to accurately judge whether there is replication of hepatitis B virus in the body and to monitor accurately after antiviral treatment. Clinically, the dynamic changes of the five serological indicators of hepatitis B and their patterns are used to monitor and monitor the condition of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N33/543G01N33/558
CPCG01N33/5764G01N33/54333G01N33/54346G01N33/558G01N2446/90
Inventor 贾鑫明王祎龙崔正权朱翰飞
Owner SUZHOU WIN BIO TECH CO LTD
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