Energy resonance transfer technology based on upconversion fluorescence for detection of exosomes

A technology of exosomes and fluorescence, which is applied in the fields of sensors, analytical chemistry and nanometers, can solve the problems of expensive instruments, time-consuming analysis, and large sample volume required to achieve high sensitivity and simplify the experimental process.

Active Publication Date: 2021-02-19
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these methods are classic, they have disadvantages such as expensive instruments, time-consuming analysis, and large sample volume.

Method used

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  • Energy resonance transfer technology based on upconversion fluorescence for detection of exosomes
  • Energy resonance transfer technology based on upconversion fluorescence for detection of exosomes
  • Energy resonance transfer technology based on upconversion fluorescence for detection of exosomes

Examples

Experimental program
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Effect test

example 1

[0044]Example 1: The preparation method of fluorescent biosensor based on fluorescence resonance energy transfer is:

[0045]1) Preparation of rare earth nitrate: rare earth oxide (RE)2O3), Including oxide (ER2O30.0766g (0.2mmol), oxidation (YB)2O30.7882G (2 mmol), yttrium oxide (Y2O31.7614G (7.8 mmol); Note: M (ER2O3) = 382.52g / mol m (YB2O3) = 394.08g / mol m (Y2O3) = 225.82 g / mol mixed in 250 ml of tri-neck flask, adding dilute nitric acid (concentrated nitric acid: water volume ratio = 1: 1), a total of 100ml, sealing film sealing, water bath magnetic stirring heating 90-100 ° C, clarify After the sealing film is opened, the sealing film is opened, and the extra nitric acid is completely volatilized to obtain a rare earth nitrate solid; 10 ml of ethanol is added after natural cooling, and the mixed rare earth nitrate ethanol solution is obtained after stirring.

[0046]The reaction equation is:

[0047]Among them, the RE represents ER, YB, and Y is 2%: 20%: 78% doped mixture.

[0048]2) ...

example 2

[0058]The fluorescence sensor prepared in Example 1 of the present invention is used to detect exosomes,Figure 4 It is a transmission electron microscopy characterization map of exosomes. The specific detection method is: mix EpCAM-1-UCNPs and EpCAM-2-TAMRA in a solution, and mix different concentrations of MDA-MB-231 exosomes (0, 1 ×103, 1×104, 1×105, 1×106, 1×107, 1×108Pieces / mL) were added to Tris hydrochloric acid buffer containing EpCAM-1-UCNPs and EpCAM-2-TAMRA, and after shaking at 28°C for 1 hour, the fluorescence intensity was measured. Such asFigure 5 As shown, with the concentration of exosomes (A to G in the figure are different concentrations of exosomes (0, 1×103, 1×104, 1×105, 1×106, 1×107, 1×108particles / mL) increases, the fluorescence intensity at 585nm gradually increases.Figure 6 It is the fitted linear relationship diagram of exosomes detection. It can be seen from the figure that the concentration of exosomes and the intensity of fluorescence at 585nm produced b...

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Abstract

The invention discloses a fluorescence energy resonance transfer based on upconversion nanomaterials and tetramethylrhodamine for the detection of exosomes, which involves rare earth-doped upconversion nanomaterials as fluorescent probe sensors, through carrying UCNPs and TAMRA Two different EpCAM protein aptamer DNA chains can recognize the EpCAM protein on the surface of exosomes and bind to the protein to form a hairpin structure, which causes the distance between UCNPs and TAMRA to be shortened to less than 10 nm, resulting in Fluorescence resonance energy transfer to generate fluorescent signals; the advantages of the sensor: rare earth-doped upconversion luminescent nanomaterials are photochemically stable due to their unique anti-Stokes luminescence properties, and have no autofluorescence; avoid the use of expensive proteins such as proteins The reagents simplify the experimental process; the method is based on the principle of fluorescence energy resonance transfer and has high sensitivity. It can be used for highly sensitive detection of exosomes derived from breast cancer cells.

Description

Technical field[0001]The invention relates to an analysis method based on the principle of fluorescence resonance energy transfer between up-conversion nanomaterials and tetramethylrhodamine, and belongs to the field of analytical chemistry and nanotechnology.[0002]The invention relates to an aptamer sensor using rare earth-doped upconversion nanomaterials as fluorescent probes, and realizes high-sensitivity detection of exosomes through changes in upconversion fluorescence intensity, and belongs to the field of sensors.Background technique[0003]Exosomes are vesicle bodies secreted by a variety of living cells, which can be used by most types of cells such as B cells, T cells, dendritic cells, macrophages, neurons, glial cells, and tumor cells. It is secreted by stem cells, etc., with a diameter of about 30-100nm. Exosomes have diverse sources and carry a large number of biological information molecules such as proteins, lipids, DNA, RNA, etc., which make them the messengers of info...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 兰建明陈敬华李春艳吴芳罗登旺方垚盛依伦
Owner FUJIAN MEDICAL UNIV
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