Fluorescence-magnetic resonance bimodal quantum dot and preparation and application method thereof

A technology of magnetic resonance and quantum dots, applied in the field of fluorescence-magnetic resonance dual-mode quantum dots and its preparation and application, can solve the problems of low selectivity of fluorine monomer, poor stability, low fluorine content, etc., and achieve an improvement Sensitivity and specificity, good hydrophilicity, small effect size

Inactive Publication Date: 2019-03-29
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the currently reported fluorinated contrast agents have several serious defects: (1) low fluorine content (<5wt%); (2) low selectivity of fluorine monomer; and (3) fluorine-based contrast agents Very few in vivo studies in animals
However, most of these encapsulated or modified particles have disadvantages such as large particle size and poor stability.

Method used

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  • Fluorescence-magnetic resonance bimodal quantum dot and preparation and application method thereof
  • Fluorescence-magnetic resonance bimodal quantum dot and preparation and application method thereof
  • Fluorescence-magnetic resonance bimodal quantum dot and preparation and application method thereof

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Experimental program
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Embodiment 1

[0048] Embodiment 1: Pdots-19F particle preparation:

[0049] This embodiment takes CNPPV, PFCE, PS-PEG-COOH and THF as examples for preparation, but the protection scope of the present invention is not limited to these selected raw materials.

[0050] Both fluorocarbons and conjugated polymers of the present invention are available from Sigma Aldrich, https: / / www.sigmaaldrich.com / united-states.html.

[0051] Specifically include the following steps:

[0052] (1) conjugated polymer and fluorocarbon are dissolved in tetrahydrofuran;

[0053] (2) Rapidly inject the solution into water in an ultrasonic water bath, and keep it in the ultrasonic water bath for at least 1 min;

[0054] (3) The tetrahydrofuran is removed after the obtained solution is heated below 100° C., that is, the conjugated polymer quantum dots wrapped with fluorocarbons are obtained.

[0055] Each raw material and its addition ratio and concentration are as follows:

[0056]

[0057] Mix CNPPV, PFCE, PS...

Embodiment 2

[0059] Embodiment 2: Pdots-19F particle surface modification:

[0060] The Pdots-19F prepared in Example 1 was cross-linked by EDC to obtain targeted Pdots-19F-biophilin nanoparticles (Pdots-19F-SA).

[0061] 1.4mL, 50ppm Pdots-19F nanoparticles were placed in a 20mL glass bottle;

[0062] 2. Add 80 μL 1M HEPES and 80 μL 5% PEG solution, and keep stirring for 5 minutes;

[0063] 3. Add 40 μL of freshly prepared, 10 mg / mL EDC solution, and keep stirring for about 10 minutes;

[0064] 4. Add 240μL, 1mg / mL bioavidin solution, and keep stirring for 3 hours;

[0065] 5. Add 80 μL of 10% BSA solution and stir for 30 minutes to terminate the reaction;

[0066] 6. Put the above solution in a 4mL ultrafiltration tube (100kDa molecular weight), and centrifuge at 3500rpm for 4 minutes;

[0067] 7. Add the concentrated solution (approximately 0.5mL) from step 6 to a size chromatography column for separation. Thereby removing free small molecules and excess protein.

Embodiment 3

[0068] Example 3: Fluorescence imaging and 19FMRI imaging of Pdots-19F-SA

[0069] 1. Culture MCF-7 breast cancer cells at 37°C.

[0070] 2. Incubate biotinylated anti-EPCAM antibody (anti-epithelial cell adhesion molecule antibody) with MCF-7 cells at room temperature for 45 minutes.

[0071] 3. Wash off excess biotinylated anti-EPCAM antibody by centrifugation (1000 rpm, 5 minutes) in 1XPBS solution.

[0072] 4. Add 1-10 ppm of Pdots-19F-SA in 1X PBS and incubate at room temperature for 45 minutes.

[0073] 5. Wash off excess Pdots-19F-SA by centrifugation (1000 RPM, 5 minutes) in 1XPBS solution.

[0074] 6. Fix the cells in formalin and start fluorescence imaging, flow cytometry and 19F NMR detection, see the results image 3 and Figure 4 .

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Abstract

The present invention discloses a fluorescence-magnetic resonance bimodal quantum dot and a preparation and an application method thereof. The method is as follows: a conjugated polymer and fluorocarbon are dissolved in tetrahydrofuran, and a solution is rapidly injected into water in an ultrasonic water bath and kept in the ultrasonic water bath for at least one minute. After the obtained solution is stirred or heated, a tetrahydrofuran solvent is removed, thereby obtaining a fluorocarbon-encapsulated conjugated polymer quantum dot which is recorded as Pdots-19F. The prepared Pdots-19F has diversity of targeted imaging, can realize combination of fluorescence imaging and 19F MRI magnetic resonance imaging, and has good sensitivity and specificity, and huge application prospects.

Description

technical field [0001] The invention belongs to the technical field of contrast agent and its preparation, and in particular relates to a fluorescence-magnetic resonance dual-mode quantum dot and its preparation and application method. Background technique [0002] Magnetic resonance (MRI), as an important means of clinical disease diagnosis, has attracted the attention of many researchers. Among them, many patients need to inject MRI contrast agent to enhance the imaging effect, so as to achieve the purpose of accurate and rapid assessment of the disease. [0003] Fluorine-containing compounds are a new class of MRI contrast agents. Compared with gadolinium contrast agents commonly used in clinical practice, they have the characteristics of low toxicity, high contrast, and quantitative assessment of disease severity. However, the currently reported fluorinated contrast agents have several serious defects: (1) low fluorine content (<5wt%); (2) low selectivity of fluorine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/12A61K49/18A61K49/00
CPCA61K49/0002A61K49/0054A61K49/0067A61K49/0093A61K49/126A61K49/186
Inventor 伍旭熊炜曾朝阳李桂源何姝仪武迎芬
Owner CENT SOUTH UNIV
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