Method for quantitatively detecting MFAP (PSI-OAm-NAPI) nano-drug carrier by fluorescence immunoassay

A nano-drug carrier and quantitative detection technology, applied in measuring devices, biomaterial analysis, instruments, etc., can solve the problems of slow metabolism of carriers, toxic and side effects of organisms, etc.

Pending Publication Date: 2018-02-23
ANHUI NORMAL UNIV
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  • Claims
  • Application Information

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Problems solved by technology

The metabolism of the carrier in the body is inseparable from the health of the organism. Excessiv

Method used

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  • Method for quantitatively detecting MFAP (PSI-OAm-NAPI) nano-drug carrier by fluorescence immunoassay
  • Method for quantitatively detecting MFAP (PSI-OAm-NAPI) nano-drug carrier by fluorescence immunoassay
  • Method for quantitatively detecting MFAP (PSI-OAm-NAPI) nano-drug carrier by fluorescence immunoassay

Examples

Experimental program
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Embodiment 1

[0105] A method for fluorescent immunoquantitative detection of MFAP nano drug carrier, comprising the following steps:

[0106] a. Preparation of MFAP whole antigen;

[0107] a-1. Take 32mL of N,N-dimethylformamide and heat it to 60°C, then add 1.6g of polysuccinimide PSI and 1.63mL of oleylamine OAm, and add 0.83mL of N-(3-amino Propyl)imidazole NAPI, and raise the temperature to 100°C for 5 hours, then cool to room temperature, add 160mL of methanol and make it uniformly precipitate under magnetic stirring, and then take the precipitate and dry it after centrifugation to prepare PSI-OAm-NAPI (MFAP);

[0108] a-2. Take 10 mg of the nanoparticles and 0.1 mg of polyethylene glycol-poly(lactic-co-glycolic acid) mPEG-PLGA, and dissolve them together in 1 mL of chloroform solution;

[0109] a-3. Add it to 10 mL of freshly prepared sodium hydroxide solution with a concentration of 0.006 mg / mL, and sonicate for 6 minutes at a power of 300 W;

[0110] a-4. Evaporate chloroform in...

Embodiment 2

[0145] Detect the concentration of the MFAP test solution of the unknown concentration of the unknown concentration.

[0146] To enrich embodiment 1, just replace the MFAP standard solution of different concentrations in step e-3 with the MFAP test solution of unknown concentration, then measure each hole on a microplate reader when the excitation wavelength is 485nm and the emission wavelength is 590nm Fluorescence intensity, calculate the average fluorescence intensity value, according to the above-mentioned standard curve can calculate the concentration of MFAP test solution.

[0147]

[0148] For standard addition recovery determination, it is only necessary to add 50 μL of the sample to be tested in each well to add 25 μL of the sample to be tested and 25 μL of the concentration of the standard, and the other conditions remain unchanged. Recovery rate = (measured value of standard addition - detection amount of sample to be tested) / concentration of standard addition ...

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Abstract

The invention provides a method for quantitatively detecting an MFAP (PSI-OAm-NAPI) nano-drug carrier by fluorescence immunoassay. Mesoporous silica nano-particles are taken as a substrate, a silicondioxide nano particle has the advantages of large specific surface area and good stability, and the loading rate of fluorescent dye is high; fluorescent dye-boron dipyrromethene dyes (BODIPY) and rhodamine B are doped, two fluorescent dyes can generate fluorescence resonance energy transfer, fluorescence intensity is improved, and quenching is reduced; a sodium periodate method coupling antibody is adopted to improve coupling efficiency, and the influence of a cross-linking agent for antibody activity is eliminated. The high specificity of the antigen antibody in immunology reaction is fully utilized, in addition, the high specificity is combined with the sensitivity of fluorescence, an immunoreaction signal is further improved, and the ultra-sensitive detection of the nano-drug carrier isrealized.

Description

technical field [0001] The invention relates to the field of quantitative detection of nanomaterials, in particular to a method for quantitatively detecting MFAP nano drug carriers by fluorescence immunoassay, capable of ultrasensitive fluorescence immunoassay detection of polysuccinimide (PSI), N-(3-aminopropyl) PSI-OAm-NAPI (MFAP) nano-drug carrier obtained through the joint action of imidazole (NAPI) and oleylamine (OAm). Background technique [0002] Nano-scale anticancer drugs have become a promising new class of cancer therapeutic agents. In addition to the anticancer drug itself, the composition of the preparation also has a drug "transporter" that plays a decisive role in drug delivery. The "transporter" mentioned here refers to nanomaterials that can be used to load drugs, that is, nano drug carriers. Nanocarriers consist of organic materials including polymers, liposomes, vesicles, polypeptides, and inorganic materials such as gold nanoparticles and quantum dots. ...

Claims

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Application Information

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IPC IPC(8): G01N33/533
CPCG01N33/533G01N2430/60
Inventor 张明翠闫希
Owner ANHUI NORMAL UNIV
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