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Preparation method of a 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 complex operation, time-consuming technical requirements, and side effects, and achieve the effects of simple preparation process, high relaxation rate, and low toxicity

Inactive Publication Date: 2020-07-28
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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  • Preparation method of a specific magnetic endoglin aptamer imaging probe system
  • Preparation method of a specific magnetic endoglin aptamer imaging probe system
  • Preparation method of a specific magnetic endoglin aptamer imaging probe system

Examples

Experimental program
Comparison scheme
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 and dissolve in 30 mL of ddH 2 Add 10 mL of PEG-2000 aqueous solution with a concentration of 50 g / L in O, stir and mix well, and add 30 mL of dilute ammonia dropwise, continue to stir quickly, and add 270 μL of H 2 O 2 After the solution turns black, continue to stir for 20 minutes; transfer the solution to an autoclave and react at a constant temperature at 160°C for 6 hours, then magnetically separate and wash 3 times and dry at 70°C to obtain magnetic Fe 3 O 4 Nanoparticles.

Embodiment 2

[0033] Magnetic Chitosan Nanoparticles Fe 3 O 4 @CMCS preparation:

[0034] Weigh 0.4 g of carboxymethyl chitosan CMCS in 20 mL ddH 2 In O, filter after stirring to dissolve completely, and the filtrate is ready for use; add 0.4 g of prepared Fe into the filtrate 3 O 4 Magnetic nanoparticles, ultrasonic for 5 min, get uniformly dispersed Fe 3 O 4 Magnetic nanoparticle dispersion; add 58.2 mL liquid paraffin oil, 1.8 mL span-80 into a 250 mL three-necked bottle; then add the evenly dispersed Fe while stirring 3 O 4 Magnetic nano-particle dispersion (water phase), the mixture is sonicated for 30 min to make it uniformly dispersed, and the reaction is fully stirred for 2 h; 2 mL of 25% glutaraldehyde solution is added, and the reaction is continued for 1.5 h; after the reaction is complete Ether magnetic separation and washing 3 times, then acetone magnetic separation and washing 3 times; the resulting product was dried in a vacuum drying oven at 70°C, and ground to obtain magnetic ch...

Embodiment 3

[0036] Magnetic Endoglin aptamer imaging probe mEND- Fe 3 O 4 @CMCS preparation:

[0037] Magnetic chitosan nanoparticles Fe 3 O 4 @CMCS (-NH2) was dissolved in HEPES buffer pH 7.2, and 40 μL sulfo-SMCC (4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid sulfosuccinyl 4-(N-Maleimidomethyl)cyclohexane-1-carboxylic acid 3-sulfo-N-hydroxysuccinimide ester sodium salt), incubate for 2 h at 4 ℃ in a constant temperature shaker, and separate the cross-linked SMCC with a magnetic stand Magnetic particles, after removing the excess crosslinking agent, add 2-Endoglin aptamer, modify the sulfhydryl group into the crosslinked magnetic particles, and mix; incubate with a constant temperature shaker at 4 ℃ for 2 h; add 100 μL of 1% cattle Serum protein was blocked for 30 min; the nanoparticles were washed 2-3 times with 1 ml coupling buffer, and the magnetic particles were separated using a magnetic stand to obtain the magnetic Endoglin aptamer imaging probe mEND-Fe 3 O 4 @CMCS.

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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, and specifically relates 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 approximately 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 related to the proliferation of endothelial cells. Related and can be induced by hypoxic environment. Nucleic acid aptamer (aptamer) is a type of nucleic acid molecule with high specificity and high affinity. It is a small molecule DNA or RNA that is screened from a random oligonucleotide library through an exponential enrichment system (SELEX). It has high affinity and specificity to the target, and can be used for disease diagnosis and targeted therapy. Magnetic nanomaterials have magnetic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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