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Multifunctional nanoprobe for targeting SERS (surface enhanced Raman scattering) imaging of tumor cells and preparation method thereof

A nano-probe, tumor cell technology, applied in the field of nano-probes, can solve the problem of a small number of targeted nano-probes, and achieve the effect of improving biocompatibility, simple structure, and high photothermal performance

Inactive Publication Date: 2016-05-25
NANJING UNIV OF POSTS & TELECOMM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to overcome the relatively small number of targeted nanoprobes that integrate SERS imaging and photothermal therapy, the present invention provides a multifunctional nanoprobe for tumor cell-targeted SERS imaging and a preparation method thereof. Using star-shaped gold nanoparticles as the main body of probe construction, and then modifying Raman molecules (mercaptobenzoic acid) on the surface of gold nanoparticles, and then further binding to a polypeptide (RGD) that can specifically bind to tumor cells, a protein that can specifically bind tumor cells was prepared. Multifunctional nanoprobes for targeted recognition, imaging and photothermal therapy of tumor cells

Method used

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  • Multifunctional nanoprobe for targeting SERS (surface enhanced Raman scattering) imaging of tumor cells and preparation method thereof
  • Multifunctional nanoprobe for targeting SERS (surface enhanced Raman scattering) imaging of tumor cells and preparation method thereof
  • Multifunctional nanoprobe for targeting SERS (surface enhanced Raman scattering) imaging of tumor cells and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0038] 1. Prepare Au seeds: first add 0.5mL5mMHAuCl to the bottle 4, then add 10mL10mMCTAC and stir for 5min, then add 0.6mL ice-water bath dropwise (1 / 20mL) to configure 10mMNaBH 4 , stirring rapidly (500~1000r / min) for 2 minutes, the color is light brown.

[0039] 2.1 mL 10 mM HAuCl 4 Add 10ml 200mMCTAC and stir for 2min, dilute the Au seeds prepared in the previous step by 100 times, add 0.05mL and stir for 2min, add 0.5mL0.3mMAA and stir for 2min, then put it in a temperature-controlled box at 15°C to grow and mature for 3h, and finally wash it by centrifugation The nanoparticles were redispersed and the volume was adjusted to 2 mL.

[0040] 3. Star-shaped Au linked with 4MBA, add 50μL, 200μL, 400μL of 1mM 4-MBA ethanol solution respectively, put it in a shaker at 25°C and react at 200r / min for 20h. 3000r / min, 6min centrifugal washing twice.

[0041] 4. To activate the carboxyl terminal of 4MBA, add 24 μL of 100 mM EDC and 59.4 μL of 100 mM NHS to react on a shaker for...

Embodiment 2

[0043] To test the photothermal properties of water and Au, set the 785nm laser to 0.4w, the distance from the laser to the 96-well plate is 17mm, and the volume of Au is set to 2mL, and 200μl of Au particle solution is added for each test. Under 785nm laser irradiation, the temperature was recorded with a handheld thermometer every 1 min, and the photothermal properties of star-shaped gold nanoparticles were tested. according to image 3 , the temperature of star-shaped gold nanoparticle solution increased by 16.4°C when the laser was irradiated for 10 min.

Embodiment 3

[0045] To test the effect of SERS probe on photothermal therapy of A549 lung cancer cells, A549 lung cancer cells (RGD targeting) were seeded in 6-well cell culture plates, and the buffer solutions contained 0.5mg / mL, 0.2mg / mL, 0.1mg / mLAu respectively -4MBA-RGD and Au-4MBA were incubated for 4 hours, the medium and colloidal particles were sucked off, washed three times with PBS, and irradiated with 785nm laser near infrared laser for 6 minutes. Wash with PBS stained with Calcein-AM (only for live cells) and PI (for dead cells), and observe the cell viability under an inverted fluorescent microscope. The result is as Figure 7 , under the same laser irradiation conditions, the higher the concentration of SERS probe input, the denser the staining of dead cells by PI, indicating the better effect of photothermal therapy.

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Abstract

The invention discloses a multifunctional nanoprobe for targeting SERS (surface enhanced Raman scattering) imaging of tumor cells and a preparation method thereof. The probe comprises gold nanoparticles, thiosalicylic acid, polypeptide and bovine serum albumin sequentially from inside to outside, the gold nanoparticles are wrapped with a layer of thiosalicylic acid molecules and then connected with the polypeptide through an amido bond, and finally the surface is wrapped with a layer of the bovine serum albumin to enclose the exposed area of the surface of the probe to obtain the nanoprobe; the gold nanoparticles are star-like gold nanoparticles. The preparation method includes: firstly, preparing the star-like gold nanoparticles high in near-infrared absorption performance, secondly, modifying the surfaces of the gold nanoparticles with Raman molecules, and thirdly, further binding the polypeptide capable of specifically binding the tumor cells. The probe integrates multiple functions, has targeting SERS imaging performance and a photothermal therapy function, can effectively recognize the tumor cells in a directional manner, performs tumor imaging, and utilizes the photothermal therapy to kill the tumor cells.

Description

technical field [0001] The invention relates to the technical field of nanoprobes, in particular to a multifunctional nanoprobe for tumor cell-targeted SERS imaging and a preparation method thereof. Background technique [0002] The application of nanotechnology in medicine is currently a rapidly developing field. Many nanomaterials with unique structural, optical, electrical, magnetic, and catalytic properties have been explored for tumor imaging, diagnosis, and therapy. Surgical resection of cancer cells is an efficient method of cancer treatment, but it has significant side effects, and it is still a challenge to treat primary tumors located in important sites or inaccessible tissues. And photothermal therapy is to raise the temperature of cancer cell tissue to 40-43 ℃ or higher. When the temperature is higher than 43°C, the protein denatures and the cell membrane is damaged. In this case, the tumor tissue will be ablated. Photothermal therapy is a minimally invasive t...

Claims

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Application Information

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IPC IPC(8): A61K41/00A61K47/48A61K49/00A61P35/00
CPCA61K41/0052A61K49/0013
Inventor 宋春元汪联辉陈文蔷窦艳霞
Owner NANJING UNIV OF POSTS & TELECOMM
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