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Near-infrared Ag2S nanometer assembly probe, and preparation method and applications thereof

A nano-assembly and near-infrared technology, applied in the field of biological analysis, to achieve high sensitivity and good selectivity

Active Publication Date: 2018-11-30
EAST CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, the development of NIR fluorescent probes for highly sensitive capture and detection of CTCs is still highly demanded and challenging.

Method used

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  • Near-infrared Ag2S nanometer assembly probe, and preparation method and applications thereof
  • Near-infrared Ag2S nanometer assembly probe, and preparation method and applications thereof
  • Near-infrared Ag2S nanometer assembly probe, and preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: DNA-Ag 2 S and Ag 2 Preparation of S assemblies

[0066] DNA-Ag 2 Preparation of S:

[0067] According to our previous method, 5 mM AgNO 3 and 25mM 3-MPA were dissolved in 30mL of deoxygenated deionized water, and the reaction was stirred at 90°C for 12h under the condition of pH=7.5 to prepare Ag 2 S. On this basis, we successfully synthesized DNA-Ag by a one-step method 2 S.

[0068] In short, 100 μM DNA of thiosulfate was added to 400 μL of quantum dot reaction precursor solution, and the reaction was stirred at 90 °C for 12 h to obtain DNA functionalized Ag 2 S. The obtained DNA-Ag was then washed using an ultrafiltration tube with a molecular weight cut-off of 3000 Da 2 S and store at 4 °C.

[0069] NIR Ag 2 Preparation of S nanoassembly probes:

[0070] Ag for isolation and detection of CTCs 2 S assembly and Fe 3 o 4 -Assembly of EpCAM, DNA-Ag 2 S and aptamers hybridize to H1 and H2 to obtain HCR under the action of the initiator to obta...

Embodiment 2

[0076] Example 2: Ag 2 Imaging sensitivity and stability investigation of S assembly:

[0077] Human breast cancer cells (MCF-7) were cultured in high-sugar DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin, and different forms of near-infrared Ag obtained during the experiment 2 The S assembly probes were incubated with well-grown MCF-7 and imaged using confocal microscopy.

[0078] Such as image 3 As shown in (A) and (B), in the data analysis results of the imaged images and their intensity after imaging, the near-infrared Ag 2 The imaging signal of the S assembly probe was significantly higher than that of DNA-Ag at the same concentration 2 S (about 2.4 times), while DNA-Ag 2 The imaging signal of S is also higher than that of Ag 2 S, which demonstrates aptamer modification and assembly of NIR Ag 2 S assembly probes can significantly increase the sensitivity of imaging.

[0079] The present invention prepares near-infrared Ag with differen...

Embodiment 3

[0082] Example 3: Fe 3 o 4 Preparation of Magnetic Nanoparticles and Coupling with Antibody EpCAM

[0083] First, 8.1g FeCl 3 ·6H 2 After O was dissolved in 142.5 mL of deionized water, it was transferred to a three-necked flask and heated to 70 °C with stirring. Then, weigh 4.4g FeCl 2 4H 2 O was dissolved in 10 mL of deionized water, filtered, and 7.5 mL was added to a three-neck flask. Under vigorous stirring, 18 mL of concentrated ammonia water with a mass fraction of 25% was quickly added, and after 1 min, 4.66 g of oleic acid was added dropwise, and stirring was continued at 70° C. for 1 h. After the reaction was completed, a dark sol was obtained, and the resulting precipitate was separated from the reaction system by applying an external magnetic field. Wash twice with ethanol to remove excess oleic acid and rinse with deionized water to pH=7. Then add 160 mL of KMnO with a concentration of 10 mg / mL 4 solution, and ultrasonically treated with an ultrasonic cle...

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Abstract

The invention provides a near-infrared Ag2S nanometer assembly probe. The near-infrared Ag2S nanometer assembly probe comprises a plurality of monomers M1 and M2 which are assembled based on HCR reaction; the monomer M1 is prepared through assembling of DNA-Ag2S nanodot and nucleic acid sequence such as H1 represented by SEQ ID NO.2, wherein the DNA sequence is represented by SEQ ID NO.1; the monomer M2 is prepared through assembling of an aptamer such as the aptamer represented by SEQ ID NO.3 and nucleic acid sequence such as H2 represented by SEQ ID NO.4. The invention also provides a preparation method of the near-infrared Ag2S nanometer assembly probe. The raw materials are easily available; the conditions are mild; normal temperature reaction is adopted; no heating is needed; productquality is stable. The invention also provides application method and applications of the near-infrared Ag2S nanometer assembly probe in detection of circulating tumor cells. The near-infrared Ag2S nanometer assembly probe is used in detection of circulating tumor cells, is high in specific identification capacity and sensitivity, can be used for further analysis of target cells in whole blood, and is promising in application prospect in cancer clinical diagnosis, postoperative prevention, and treatment.

Description

technical field [0001] The invention belongs to the field of biological analysis, and in particular relates to a near-infrared Ag 2 S nanometer assembly probe, its preparation method and its application in detection of circulating tumor cells. Background technique [0002] Circulating tumor cells (CTCs) are a general term for tumor cells present in peripheral blood, and their expression levels in blood are closely related to the prognosis and survival of cancer patients. The detection of CTCs is of great significance for the early diagnosis of metastases, cancer prognosis, treatment monitoring and metastasis diagnosis. But the detection and analysis of CTCs is not easy because there is no clear distinction between CTCs and other blood cells. In addition, the number of CTCs in peripheral blood is very small, usually in the range of 10 9 Only one in every white blood cell. Therefore, highly efficient isolation and detection of CTCs from whole blood is a challenging subject...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N21/85
CPCG01N21/64G01N21/8507G01N2021/656
Inventor 鲜跃仲张翠玲丁彩萍
Owner EAST CHINA NORMAL UNIVERSITY
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