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Method used for fluorescent immunoadsorption measurement of ultra trace high-molecular nanometer drug carrier based on magnetic fluorescence probe labelling

A nano-drug carrier and fluorescent probe technology, which is applied in the field of functionalization and quantitative detection of nano-materials, can solve problems such as poor photobleaching resistance, immature detection technology, and large sample damage, and achieve enhanced magnetic separation effects and improved Photobleaching resistance, uniform particle size distribution effect

Active Publication Date: 2018-11-30
ANHUI NORMAL UNIV
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  • Abstract
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Problems solved by technology

Fluorescent reagents are usually used as imaging agents for nano-drug carriers, but due to the poor photobleaching resistance of various fluorescent reagents encapsulated in drug carriers, some fluorescent imaging agents may even interact with drugs, resulting in changes in drug efficacy.
Therefore, it is impossible to achieve long-term real-time dynamic detection by using fluorescent imaging agents to detect single nano-drug carriers.
[0008] However, the commonly used quantitative analysis methods for nanomaterials are mainly based on expensive analytical instruments.
Moreover, the reagents used in the test have certain toxicity and great damage to the sample, the stability and sensitivity often fail to meet the requirements, and the detection technology is still not mature enough.
Therefore, these methods are unable to achieve real-time accurate quantitative detection and analysis of nanomaterials.

Method used

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  • Method used for fluorescent immunoadsorption measurement of ultra trace high-molecular nanometer drug carrier based on magnetic fluorescence probe labelling

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preparation example Construction

[0051] The preparation method of each solution that the present invention relates to is:

[0052] PBS solution (0.01mol / L pH=7.4): weigh 8.0g of NaCl, 0.1g of KCl, NaH 2 PO 4 2H 2 O0.106g, Na 2 HPO 4 12H 2 O 3.34g was dissolved in distilled water and the volume was adjusted to 1000mL.

[0053] Carbonate buffer solution CD (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.

[0054] PBST solution (0.01mol / L pH=7.4): add 500 μL Tween-20 to 1000 mL PBS, and mix well.

[0055] Coating buffer fe (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.

[0056] Acetic acid buffer: Weigh 84.25g of sodium acetate, dissolve it in water, add 100ml of acetic acid, and dilute to 2500ml with water to obtain (PH=4.2) acetic acid sodium acetate buffer solution.

[0057] 1wt% casein solution: it is a blocking solution, weigh 0.01g...

Embodiment 1

[0059] A method for measuring ultra-trace macromolecular nano drug carriers based on fluorescent immunosorbent labeling with magnetic fluorescent probes, comprising the following steps:

[0060] a. Preparation of PSI OAm Hydrolysis solution:

[0061] 20 ~ 60mg of PSI OAm Dissolve in 1-5mL chloroform solution, after dissolving, add the above solution into 10-16mL sodium hydroxide solution with a concentration of 0.001-0.008mg / mL, ultrasonic for 10-30min, power 300-600W, then magnetically stir for 20 ~50min, evaporate the solution at 40~60℃ to remove chloroform, centrifuge at 12000~20000r / min for 10~15min, wash 3 times with PBS with pH=7.4, take the precipitate and disperse it in 0.5~2mL PBS with pH=7.4 buffer to obtain the hydrolyzed PSI OAm -COO - solution.

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Abstract

The invention provides a method used for fluorescent immunoadsorption measurement of an ultra trace high-molecular nanometer drug carrier based on magnetic fluorescence probe labelling. The detectionsensitivity of the method is increased based on the easy separation performance and the excellent fluorescence performance of the magnetic fluorescence probe. According to the method, ultra trace detection analysis on oleylamine grafted polysuccinimide (PSIOAm) high molecular nanometer particles is carried out through following steps: a PSIOAm coated antigen and a magnetic fluorescence probe labelled antiPSIOAm antibody are prepared, and combination with the specificity of antigen antibody reaction is adopted. Compared with the prior art, the method is simple in operation, low in background, high in sensitivity and specificity, and is capable of realizing high flux accurate targeting nondestructive test.

Description

technical field [0001] The invention relates to the functionalization of magnetic fluorescent probes and the quantitative detection of nanomaterials, in particular to a method for measuring ultra-trace polymer nano drug carriers based on fluorescent immunoadsorption labeled with magnetic fluorescent probes. Background technique [0002] According to the 2017 China Cancer Hospital Market Research and Development Trend Forecast Report released by China Industry Research Network, at present, the incidence of cancer in my country is increasing at a rate of 3%-5% every year, and the average annual cancer medical expenses are more than 150 billion yuan. The increasing incidence and death rate of malignant tumors will inevitably bring about the demand for tumor treatment services. Therefore, it is increasingly necessary to study the development and supply and demand of efficient anti-tumor drugs. [0003] At present, the clinical treatment of cancer mainly uses drugs combined with c...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/533G01N33/543G01N21/64
CPCG01N21/6428G01N33/531G01N33/533G01N33/54346
Inventor 张明翠李磊
Owner ANHUI NORMAL UNIV