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Method for determining ultratrace oleylamine grafted polysuccinimide based on carbon dot marked fluorometric immunosorbent method

A polysuccinimide and fluorescent immunological technology, which is applied in measurement devices, biological tests, material inspection products, etc., can solve the problems of immature detection technology, large sample destructiveness, toxicity, etc., and achieve a simple green preparation method, anti- Strong photobleaching, easy-to-use effect

Inactive Publication Date: 2018-04-24
ANHUI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The commonly used quantitative analysis methods for nanomaterials include inductively coupled plasma mass spectrometry, high performance liquid chromatography-inductively coupled plasma mass spectrometry, and gas chromatography-inductively coupled plasma mass spectrometry. The reagents used are somewhat toxic and destructive to the sample, the stability and sensitivity often fail to meet the requirements, and the detection technology is still not mature enough
Therefore, none of these methods can achieve accurate quantitative detection and non-destructive analysis of nanomaterials.

Method used

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  • Method for determining ultratrace oleylamine grafted polysuccinimide based on carbon dot marked fluorometric immunosorbent method

Examples

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preparation example Construction

[0045] The preparation method of each solution that the present invention relates to is:

[0046] PBS solution (0.01mol / L pH=7.4): 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.

[0047] 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.

[0048] PBST solution (0.01mol / L pH=7.4): add 500 μL Tween-20 to 1000 mL PBS, and mix well.

[0049] 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.

[0050] 1wt% casein solution: it is a blocking solution, weigh 0.01g of casein and dissolve it in 1mL of PBS, and mix well.

Embodiment 1

[0052] A method for measuring ultra-trace amounts of oleylamine grafted polysuccinimide based on carbon dot-labeled fluorescent immunosorbent assay, the method comprises the following steps:

[0053] a. Preparation of PSI OAm Coating antigen and PSI OAm The immunogen specifically comprises the following steps:

[0054] a-1. Hydrolyzed PSI OAm , to get the PSI after hydrolysis OAm solution;

[0055] With reference to the method in Nano Research, 2015, 8(6):1932-1943, a chloroform solution of oleylamine-grafted polysuccinimide was prepared, and the steps were as follows: take 32mL N, N-dimethylformaldehyde Heat the amide to 90°C, add 1.6g polysuccinimide and 2.17mL oleylamine, keep heating at 100°C for 5 hours, finally add 80mL methanol to precipitate, centrifuge, weigh the precipitate and disperse in 2-10mL chloroform , the solution concentration was adjusted to 60 mg / mL to obtain a chloroform solution of oleylamine grafted polysuccinimide.

[0056] Take 0.5mL of this sol...

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Abstract

The invention discloses a method for determining ultratrace oleylamine grafted polysuccinimide based on a carbon dot marked fluorometric immunosorbent method. The method comprises the following steps:preparing PSIOAm coating antigen and immunogen, injecting the immunogen into an animal body to obtain a high-specificity antibody, then preparing C-dots marked PSIOAm antibody into a fluorescent antibody, and then establishing carbon dot fluorometric immunosorbent to accurately determine the content of PSIOAm. By combining the specificity of the antigen and antibody reaction and the carbon dot with high fluorescent quantum yield, the PSIOAm nano particles are quantitatively detected and analyzed. A linear range of the detection method is 5*10<-4> to 5*10<2>ng / mL, and the detection limit can reach 0.15 pg / mL.

Description

technical field [0001] The invention relates to the quantitative detection of nanomaterials, in particular to a method for measuring ultra-trace oleylamine grafted polysuccinimide based on carbon dot-labeled fluorescence immunoabsorption. Background technique [0002] According to statistics, about 8 million people die of cancer every year in the world, and nearly 14 million people face the threat of cancer death. Cancer has also become the number one cause of death, and it is an "incurable disease" in many people's minds. Since early cancers usually have no symptoms and tumors are often discovered at an advanced stage, it is extremely important to prevent cancer from occurring. At present, chemotherapy and radiotherapy are mainly used in the clinical treatment of cancer, but the current chemotherapy drugs have poor water solubility and no targeting. They will kill normal cells while killing cancer cells, causing serious side effects. It hinders the development and applica...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/533
CPCG01N33/53G01N33/533
Inventor 张明翠李磊
Owner ANHUI NORMAL UNIV
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