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A method for detecting folic acid functionalized silicon dioxide targeted nano drug carrier
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A nano-drug carrier, silica technology, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as poor stability, weak fluorescence signal, and low sensitivity
Active Publication Date: 2019-06-04
ANHUI NORMAL UNIV
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Fluorescent dyes are often used as probes for fluorescence detection and analysis methods, and their performance directly determines the feasibility and sensitivity of the detection method. Traditional fluorescent dyes such as fluorescein, rhodamine, cyanine, and rare earth complex fluorescent dyes, etc. There are inherent defects such as weak fluorescent signal, poor stability, low sensitivity, etc., which limit their wide application
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[0074] The preparation method of each solution that the present invention relates to is:
[0075] Phosphate buffered saline (PBS, 0.01mol / L pH=7.4): Weigh 8.0g of NaCl, 0.1g of KCl, NaH 2 PO 4 2H 2 0 0.29g, Na 2 HPO 4 12H 2 O 2.96g was dissolved in distilled water and the volume was adjusted to 1000mL.
[0076] PBST solution (0.01mol / L pH=7.4): add 500 μL Tween-20 to 1000 mL PBS, and mix well.
[0077] 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.
[0078] 1.5wt% casein solution: it is a blocking solution, weigh 0.15g casein and dissolve it in 10mL PBS, mix well.
[0085] Mix 380 μL tetraethyl orthosilicate TEOS and 12 mL absolute ethanol, add 570 μL 25% ammonia water, stir at 28°C for 24 hours, then add 400 μL 3-aminopropyltriethoxysilane APTES, continue stirring at 28°C for 24 hours, The reaction solution was centrifuged (10000rpm / min, 20min), the precipitate was washed with absolute ethanol, and dried at 28°C for 24h to obtain SiO 2 -NH 2 Nanoparticles;
[0087] Dissolve 57.5mg of folic acid FA in 2mL of MES buffer, then add 40.3mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimidehydrochloride EDAC ...
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Abstract
The invention provides a method for detecting a folate functionalized silica targeted nano-drug carrier. The method comprises the following steps: preparing a SiO2-FO envelope antigen and an immunogen, and preparing an anti-SiO2-FO polyclonal antibody; cross-linking the prepared BODIPY-PFNPs and the anti-SiO2-FO antibody; by taking SiO2-FO-OVA as an envelope antigen and taking a BODIPY-PFNPs labeled anti-SiO2-FO antibody as a primary antibody, establishing direct competitive fluorescenceimmunoassay to perform quantitative determination on SiO2-FO; and by taking a logarithm of the SiO2-FO standard concentration as a horizontal coordinate and taking a fluorescence intensity level as a vertical coordinate, drawing a standard curve, thereby quantitatively determining the concentration of theSiO2-FO. Compared with the prior art, the method disclosed by the invention has the characteristics of being simple in operation, high in sensitivity, high in specificity and capable of realizing highthroughput test, and the like.
Description
technical field [0001] The present invention relates to the quantitative detection of nanomaterials, in particular to a method for detecting folic acid-functionalized silicon dioxide targeting nano-medicine carriers, using novel polymer nanoparticles (BODIPY-PFNPs) wrapped in BODIPY to mark the fluorescent immunoassay method to determine the function of folic acidsilicon dioxide (SiO 2 -FO) a method for targeting nano drug carriers. Background technique [0002] Tumor is a kind of disease with complex mechanism of action caused by genetic instability and accumulation of molecular diversity mutations. It is one of the fastest-spreading diseases in today's society, with more than 10 million new cases every year. People have made great efforts to reduce the morbidity and mortality of tumors, but there is still a lack of effective treatments technically. The main reason is that there are many problems in traditional chemotherapy, including poor solubility of chemotherapeutic ...
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