Method for preparing specific magnetic Endoglin aptamer imaging probe system

An imaging probe and aptamer technology, applied in the field of biomedical engineering, can solve the problems of time-consuming technical requirements, complicated operation, and side effects, and achieve the effects of high relaxation rate, simple preparation process and low toxicity

Inactive Publication Date: 2017-08-11
GUANGXI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The invention patent with the publication number CN201410209779.8 discloses a targeted anti-liver cancer nanoparticle and its preparation method. This method is complex, time-consuming, technically demanding and has side effects
Preparation of magnetic mEND-Fe modified by Endoglin aptamer 3 o 4 @CMCS nanoprobe, the method of this probe that can specifically bind to neoplastic vascular endothelial cells of liver cancer has not been reported

Method used

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  • Method for preparing specific magnetic Endoglin aptamer imaging probe system
  • Method for preparing specific magnetic Endoglin aptamer imaging probe system
  • Method for preparing specific magnetic Endoglin aptamer imaging probe system

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Preparation of magnetic Fe by hydrothermal method 3 o 4 Nanoparticles:

[0031] Weigh 2.502 g (0.009 mol) of ferrous sulfate heptahydrate, dissolve in 30 mL of ddH 2 O, and add 10 mL of PEG-2000 aqueous solution with a concentration of 50 g / L, stir and mix well, and add 30 mL of dilute ammonia water drop by drop, continue to stir rapidly, and add 270 μL H2 o 2 solution, until the solution turns black, continue to stir for 20 min; transfer the solution to a high-pressure reactor, react at a constant temperature of 160 °C for 6 h, magnetically separate and wash 3 times, and dry at 70 °C to obtain the magnetic Fe 3 o 4 Nanoparticles.

Embodiment 2

[0033] Magnetic Chitosan Nanoparticle Fe 3 o 4 Preparation of @CMCS:

[0034] Weigh carboxymethyl chitosan CMCS 0.4 g in 20 mL ddH 2 In O, filter after stirring and dissolving completely, and the filtrate is set aside; add 0.4 g of Fe prepared in the filtrate 3 o 4 Magnetic nanoparticles, sonicated for 5 min to obtain evenly dispersed Fe 3 o 4 Magnetic nanoparticle dispersion; add 58.2 mL of liquid paraffin oil and 1.8 mL of span-80 to a 250 mL three-necked bottle; then add the evenly dispersed Fe 3 o 4 Magnetic nanoparticle dispersion (aqueous phase), the mixed solution was ultrasonicated for 30 min to disperse evenly, fully stirred for 2 h; add 2 mL of 25% glutaraldehyde solution, and continue stirring for 1.5 h; after the reaction was complete, use petroleum Ether magnetic separation and washing 3 times, and then magnetic separation and washing with acetone 3 times; the resulting product was dried in a vacuum oven at 70°C, and ground to obtain magnetic chitosan nanop...

Embodiment 3

[0036] Magnetic Endoglin aptamer imaging probe mEND-Fe 3 o 4 Preparation of @CMCS:

[0037] Magnetic chitosan nanoparticles Fe 3 o 4 @CMCS(-NH2) was dissolved in HEPES buffer at pH 7.2, and 40 μL of sulfo-SMCC (4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid sulfosuccinyl Imino ester sodium salt (4-(N-Maleimidomethyl)cyclohexane-1-carboxylic acid 3-sulfo-N-hydroxysuccinimideester sodium salt)), incubated at 4 °C for 2 h on a constant temperature shaker, and separated the magnetic cross-linked SMCC with a magnetic stand. Particles, after removing excess cross-linking agent, add 2-Endoglin aptamer, modify sulfhydryl group into the cross-linked magnetic particles, mix well; use temperature constant temperature shaker to incubate at 4 ℃ for 2 h; add 100 μL of 1 % bovine serum The protein was blocked for 30 min; the nanoparticles were washed 2-3 times with 1 ml coupling buffer, and the obtained magnetic particles were separated using a magnetic frame to obtain the magnetic E...

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Abstract

The invention belongs to the field of biomedical engineering, and particularly relates to a method for preparing a specific magnetic Endoglin aptamer imaging probe system (mEND-Fe3O4@CMCS). Magnetic Fe3O4 nano-particles are prepared with the hydrothermal method, magnetic chitosan nano-particles Fe3O4@CMCS are prepared with glutaraldehyde as the cross-linking agent by means of the emulsification and cross-linking method, and then the surface of Fe3O4@CMCS is modified with an Endoglin aptamer by means of the cross-linking action of Sulfo-SMCC, so that the magnetic Endoglin aptamer imaging probe mEND-Fe3O4@CMCS is obtained. The magnetic Endoglin aptamer imaging probe mEND-Fe3O4@CMCS prepared with the method can effectively target mouse tumor angiogenesis endothelial cell (mTEC) surface CD105 molecules, has hepatic transplantable tumor mouse MRI imaging capability, and has the advantages that sensitivity is high, T2 imaging efficiency is high, the MRI contrast effect is good, toxicity is low, the relaxation rate is high, biocompatibility is high, and liver and spleen targeting can be realized.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, in particular to a targeted specific magnetic Endoglin aptamer imaging probe system (mEND-Fe 3 o 4 @CMCS) method of preparation. Background technique [0002] Endoglin (CD105) is a cell membrane glycoprotein with a molecular weight of about 68 kd. It is one of the components of the transforming growth factor-β (TGF-β) receptor complex. It can exist independently on the cell surface and is associated with endothelial cell proliferation. Related proteins that are induced by hypoxic environments. Nucleic acid aptamer (aptamer) is a kind of nucleic acid molecule with high specificity and high affinity. It is a small molecule DNA or RNA obtained by screening from a random oligonucleotide library through an exponential enrichment system (SELEX), so it It has high affinity and specificity to the target and can be used for disease diagnosis and targeted therapy. Magnetic nanomaterials have magne...

Claims

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Application Information

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IPC IPC(8): A61K49/18A61K49/08
CPCA61K49/1863
Inventor 赵永祥周素芳黄盈盈黄勇
Owner GUANGXI MEDICAL UNIVERSITY
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