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Nuclear magnetism-optical dual-mode imaging biconical DNA nanoprobe, and preparation method and application thereof

A dual-mode imaging and nanoprobe technology, applied in the field of nanomedicine, can solve the problems of increasing toxicity, difficulty in early diagnosis of tumors, and improving contrast agents.

Active Publication Date: 2019-09-20
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the non-specificity of gadolinium contrast agent and the low sensitivity of MRI, early diagnosis of tumors is difficult
Increasing the dose of contrast media also increases the risk of gadolinium toxic effects in the body

Method used

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  • Nuclear magnetism-optical dual-mode imaging biconical DNA nanoprobe, and preparation method and application thereof
  • Nuclear magnetism-optical dual-mode imaging biconical DNA nanoprobe, and preparation method and application thereof
  • Nuclear magnetism-optical dual-mode imaging biconical DNA nanoprobe, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1 Preparation of biconical DNA nanoprobes for NMR-optical dual-modal imaging

[0102] In this embodiment, the following method is used to prepare the NMR-optical dual-mode imaging biconical DNA nanoprobe, including the following steps:

[0103] 1. Preparation of DNA-DOTA(Gd) MRI contrast agent

[0104] The nuclear magnetic resonance imaging contrast agent is prepared by two-step organic chemical reactions. The specific steps are as follows: the first step, the 5' end amino modification, the 3' end Cy3 modified single-stranded DNA was dissolved in deionized water, and the concentration was adjusted to 200 μM; DOTA-NCS was dissolved in 80% DMSO, and the concentration was adjusted to 200 mM; Strand DNA and DOTA-NCS were mixed in 0.1M HEPES (pH=9, containing 300mM MgCl 2 ) buffer solution, add a magnet to stir, and react in a water bath at 37°C for 24h. This step reaction is the coupling reaction of amino group and isothiocyanate. In the second step, the obtaine...

Embodiment 2

[0126] Example 2 Preparation of biconical DNA nanoprobes for NMR-optical dual-modal imaging

[0127] The difference between this example and Example 1 is that the nuclear magnetic resonance imaging contrast agent selected in the process of synthesizing the nuclear magnetic-optical dual-modal imaging biconical DNA nanoprobe is DNA-DTPA (Gd), that is, DTPA and gadolinium are selected. Chelation. In addition, the selection of other raw materials, the preparation method and the reaction conditions are the same as those in Example 1, so as to prepare a biconical DNA nanoprobe for NMR-optical dual-mode imaging.

Embodiment 3

[0128] Example 3 Live Cell Uptake of Biconical DNA Nanoprobes by NMR-Optical Dual Mode Imaging

[0129] In this embodiment, the following methods are used to investigate the uptake of living cells by NMR-optical dual-modal imaging biconical DNA nanoprobes, as follows:

[0130] Due to the limitation of the excitation wavelength of the laser confocal scanning microscope (CLSM), the Dylight800 fluorescent molecule modified on the DNA probe was replaced by Cy5.5. MDA-MB-231 cells were cultured in a medium containing 10% fetal bovine serum and 1% double antibody (the double antibody was a mixture containing penicillin (10,000IU) and streptomycin (10,000μg / mL) at 100 times the working concentration) High-glucose DMEM complete medium, the culture environment is 37 ° C and 5% CO 2 . When the cells grow to a confluence of about 80%, the cells are digested and re-seeded in 35mm 2 In a confocal small dish, culture overnight for 12 hours. Replace the culture medium with DNA-DOTA (Gd),...

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Abstract

The invention provides a nuclear magnetism-optical dual-mode imaging biconical DNA nanoprobe, and a preparation method and an application thereof. The dual-mode imaging DNA probe is prepared from a DNA nanostructure, a nucleic acid aptamer, a chemical fluorescent molecule and a nuclear magnetism resonance imaging contrast agent. The DNA probe provided by the invention improves the stability of the nuclear magnetism resonance imaging contrast agent and the chemical fluorescent molecule in body circulation, reduces the potential biological toxicity of gadolinium ions, and improves the targeting ability of tumors by using the nucleic acid adapter, and maintains the T1 relaxation efficiency equivalent to that of a clinical nuclear magnetism resonance contrast agent when the gadolinium content is only 1 / 20 of the clinical dose. With the synergistic effect of magnetic resonance imaging and fluorescence imaging, the DNA probe achieves higher resolution ratio and sensitivity of tumor imaging, and provides powerful technical means for early diagnosis of tumors.

Description

technical field [0001] The invention belongs to the field of nanomedicine, and in particular relates to a nuclear magnetic-optical dual-mode imaging biconical DNA nanoprobe and its preparation method and application. Background technique [0002] Magnetic resonance imaging (MRI) is widely used in tumor diagnosis due to its advantages of high imaging resolution, low interference, and imaging in any direction and plane. MRI contrast agents refer to preparations that shorten the relaxation time of human tissues such as T1 and T2 to increase the contrast of images, and preparations using paramagnetic or superparamagnetic substances have become the mainstream. The contrast agent is used to amplify all or part of the signal of the desired tissue, or to weaken the signal of the surrounding tissue, so as to amplify the contrast between light and dark. Gadolinium (Gd) is a rare earth metal element, and its atomic orbital contains seven unpaired electrons, which is the metal element ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/115A61K49/00A61K49/10A61K49/12
CPCA61K49/0002A61K49/0032A61K49/0052A61K49/0054A61K49/10A61K49/105A61K49/108A61K49/126C12N15/11C12N15/115C12N2310/16
Inventor 丁宝全王昭然宋琳琳王婷蒋乔
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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