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Method for quantitatively detecting SiO2-FO targeted nano-drug carrier on basis of direct competitionfluorescence immunoassay

A nano-drug carrier and fluorescent immunology technology, which is applied in the fields of fluorescence/phosphorescence, measuring devices, and material analysis through optical means, can solve the problems of expensive instruments, cumbersome operations, low stability and sensitivity, and achieve strong specificity, Simple operation and high sensitivity effect

Active Publication Date: 2017-10-10
UNITED POWER PHARMA TECH CO LTD
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Problems solved by technology

However, there are not many quantitative analyzes of nanomaterials. Currently, commonly used methods include inductively coupled plasma mass spectrometry (ICP-MS), high performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS) and gas chromatography-mass chromatography. Inductively coupled plasma mass spectrometry (GC-ICP-MS), however, these methods are not only cumbersome to operate, expensive instruments, but also have low stability and sensitivity, and the detection technology is still immature

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  • Method for quantitatively detecting SiO2-FO targeted nano-drug carrier on basis of direct competitionfluorescence immunoassay

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preparation example Construction

[0048] The preparation method of each solution that the present invention relates to is:

[0049] PBS solution (0.01mol / L pH=8.0): weigh 8.0g of NaCl, 0.1g of KCl, NaH 2 PO 4 2H 2 00.106g, Na 2 HPO 4 12H 2 O 3.34g was dissolved in distilled water and the volume was adjusted to 1000mL.

[0050] Carbonate buffer CB (0.5mol / L pH=9.6): weigh Na 2 CO 3 1.59g, NaHCO 3 2.94g was dissolved in distilled water and the volume was adjusted to 100mL.

[0051] PBST solution (0.01mol / L pH=8.0): add 500 μL Tween-20 to 1000 mL PBS, and mix well.

[0052] Coating buffer CB (0.05mol / L pH=9.6): weigh Na 2 CO 3 1.59g, NaHCO 3 2.94g was dissolved in distilled water and the volume was adjusted to 1000mL.

[0053] 1wt% casein solution: it is a blocking solution, weigh 0.01g of casein and dissolve it in 1mL of PBS, and mix well.

[0054] MES buffer solution (0.1M, pH=5.5): Dissolve 0.1921g MES in 10mL distilled water, adjust its pH to 5.5 with NaOH solution.

Embodiment 1

[0056] Quantitative detection of folic acid functionalized silica (SiO2) based on direct competition fluorescence immunoassay 2 -FO) a method for targeting a nano-drug carrier, the method comprising the steps of:

[0057] a. SiO 2 - Preparation of FO-coated antigens and immunogens

[0058] a-1. Weigh 10mg SiO2 2 -FO was dissolved in 1mL PBS solution, and magnetically stirred for 1h to obtain SiO with a concentration of 10mg / mL 2 -FO solution;

[0059] a-2. Weigh 10mg OVA and dissolve it in 2mL PBS, and add the SiO obtained in step a-1 while stirring 2 -FO solution, after mixing evenly, add 90 μL 25% glutaraldehyde solution dropwise, and stir for 4 hours at 25°C in the dark; put the reaction solution in a dialysis bag with a molecular weight cut-off of 8000-12000Da, dialyze it with PBS for 24 hours and collect it Get SiO 2 -FO-coated antigen with a concentration of 3.33 mg / mL, stored at 4°C until use;

[0060] a-3. Weigh 10mg BSA and dissolve it in 2mL PBS, and add the S...

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Abstract

The invention relates to a method for quantitatively detecting a SiO2-FOtargeted nano-drug carrier on the basis of direct competitionfluorescence immunoassay. SiO2-FO nano particles are subjected to quantitative detection and analysis through combination with specificity of antigen and antibody reactions and sensitivity of fluorescence. The method comprises steps as follows: a SiO2-FOenvelope antigen and animmunogen are prepared, the immunogen is injected into a body of an animal, a high-specificity antibody is obtained and marked with FITC(fluorescein isothiocyanate), a fluorescent antibody is prepared, and content of SiO2-FO is accurately measured through established direct competitionfluorescence immunoassay. The detection method is simple to operate, sensitivity and specificity are high, and high-throughput detection can be realized.

Description

technical field [0001] The invention relates to the quantitative detection of nanomaterials, in particular to a method for quantitative detection of folic acid functionalized silicon dioxide (SiO2) based on direct competition fluorescence immunoassay 2 -FO) a method for targeting nano drug carriers. Background technique [0002] The invention relates to the quantitative detection of nanomaterials, in particular to a quantitative detection of folic acid functionalized silicon dioxide (SiO2) based on direct competition fluorescence immunoassay. 2 -FO) a method for targeting nano drug carriers. Background technique [0003] Cancer is one of the diseases that seriously endanger human health. Cancer treatment is of great significance for improving the quality of life of patients, reducing the suffering of patients and reducing mortality. At present, chemotherapy is mainly used in the clinical treatment of cancer, but the current chemotherapy drugs are not targeted, and they...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/533G01N33/535G01N21/64
CPCG01N21/6402G01N21/6428G01N21/6452G01N33/533G01N33/535G01N33/574G01N2021/6441
Inventor 张明翠夏濛
Owner UNITED POWER PHARMA TECH CO LTD
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