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Near-infrared fluorescent silver nucleinate nanometer cluster as well as preparation method and applications thereof

A technology of fluorescent nucleic acid silver and silver nanoclusters, applied in biochemical equipment and methods, fluorescence/phosphorescence, chemical instruments and methods, etc., can solve the problem of lack of tumor tissue targeting

Active Publication Date: 2017-08-18
上海谱尼医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, near-infrared imaging mainly uses near-infrared fluorescent dyes and near-infrared gold quantum dots, but lacks the targeting of tumor tissues.

Method used

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  • Near-infrared fluorescent silver nucleinate nanometer cluster as well as preparation method and applications thereof
  • Near-infrared fluorescent silver nucleinate nanometer cluster as well as preparation method and applications thereof
  • Near-infrared fluorescent silver nucleinate nanometer cluster as well as preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 (the optimal preparation scheme of Trp-DNA-AgNCs@BSA): Using the DNA sequence as a template, add 6 μmol of DNA (the DNA sequence is "5'CCCACCCACCCTCCCAACAACAGAGGAG3 '") (the sequence was designed by ourselves, synthesized with a DNA synthesizer of Shanghai Bio-Shenggong Co., Ltd., purified by HPLC, and the sequence verified by mass spectrometry was 5'CCCACCCACCCTCCCAACAACAGAGGAG3'), 200μmolAgNO 3 (where DNA and Ag + The molar ratio of 1:33.33) and 150 μmol tryptophan (where tryptophan and Ag + The molar ratio is 1:1.33), mixed evenly and placed in a water bath, heated at 71°C for 2 minutes, then began to cool down, the cooling process took 2 hours, and then dropped to room temperature. Add NaBH to the reaction solution at room temperature 4 solution (wherein NaBH 4 with Ag + The molar ratio of the Trp-DNA-Ag + Reduction to Trp-DNA-Ag, the reaction solution was transferred to 4 ° C environment and kept in the dark for 2 days, and then added 3 μmol BSA (whe...

Embodiment 2

[0018] Example 2: Using the DNA sequence as a template, add 10 μmol of DNA (the DNA sequence is "5'CCCACCCACCCTCCCAACAACAGAGGAG3'") in 1 mL of sodium citrate buffer solution (pH 5.0, 10 mM) (the sequence is designed by ourselves, using Shanghai Biological Synthesized by DNA synthesizer of Sangon Company, purified by HPLC, and the sequence verified by mass spectrometry is 5'CCCACCCACCCTCCCAACAACAGAGGAG3'), 200μmol AgNO 3 (where DNA and Ag + The molar ratio of 1:20) and 200μmol tryptophan (where tryptophan and Ag + The molar ratio is 1:1), mixed evenly, placed in a water bath, heated at 71°C for 2 minutes, and then began to cool down. The cooling process took 2 hours, and then dropped to room temperature. Add NaBH to the reaction solution at room temperature 4 solution (wherein NaBH 4 with Ag + The molar ratio of the Trp-DNA-Ag + Reduction to Trp-DNA-Ag, the reaction solution was transferred to 4 ° C environment and kept in the dark for 2 days, and then added 4 μmol BSA (wh...

Embodiment 3

[0019] Example 3: Using the DNA sequence as a template, add 5 μmol of DNA (the DNA sequence is "5'CCCACCCACCCTCCCAACAACAGAGGAG3'") in 1 mL of sodium citrate buffer solution (pH 5.0, 10 mM) (the sequence is designed by ourselves, using Shanghai Biological Synthesized by DNA synthesizer of Sangon Company, purified by HPLC, and the sequence verified by mass spectrometry is 5'CCCACCCACCCTCCCAACAACAGAGGAG3'), 200μmol AgNO 3 (where DNA and Ag + The molar ratio of 1:40) and 100μmol tryptophan (where tryptophan and Ag + The molar ratio is 1:2), mixed evenly, placed in a water bath, heated at 71°C for 2 minutes, and then began to cool down. The cooling process took 2 hours, and then dropped to room temperature. Add NaBH to the reaction solution at room temperature 4 solution (wherein NaBH 4 with Ag + The molar ratio of the Trp-DNA-Ag +Reduction to Trp-DNA-Ag, the reaction solution was transferred to 4 ° C environment for 2 days in the dark, and then added 2.5 μmol BSA (wherein BSA...

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Abstract

The invention belongs to the fields of cell imaging and preparation of nanometer nucleic acid quantum dot reagents, and in particular relates to a stable near-infrared silver nucleinate nanometer cluster as well as a preparation method and applications of the stable near-infrared silver nucleinate nanometer cluster in cell imaging. The nucleotide sequence is taken as a template, in sodium citrate buffer solution, NaBH4 is adopted for reducing AgNO3, and thus the silver nucleinate nanometer particles with the near-infrared fluorescence are synthesized. The silver nucleinate nanometer particles can emit 800nm fluorescent light under the exciting wavelength being 750nm, the size of the silver nucleinate nanometer cluster is 70nm. The silver nucleinate is high in stability, and can be used for the fluorescent imaging of tumor cells.

Description

technical field [0001] The invention belongs to the field of cell imaging and preparation of nano nucleic acid quantum dot reagents, in particular to a stable near-infrared nucleic acid silver nanocluster and its method and application in cell imaging. Background technique [0002] Using DNA as a template to prepare inorganic nanomaterials and nanoclusters has become a new application of DNA. Among them, DNA-templated silver nanoclusters have potential applications in biosensing. Compared with quantum dots and organic dyes, nucleic acid silver nanoclusters (DNA-AgNCs) have the advantages of easy synthesis, excellent optical properties and tunable emission. However, most DNA-AgNCs have a short fluorescence lifetime, the fluorescence intensity decreases with time, and is quenched in a short time, which limits its application in biological probes (Zhou Z X and Dong S J, Protein-DNA interactions: A novel approach to improve the Fluorescent stability of DNA / Agnanoclusters, Nano...

Claims

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Application Information

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IPC IPC(8): C12N15/11G01N21/64C09K11/06
CPCC09K11/06C09K2211/181C12N15/11C12N2310/10G01N21/6486C12N2330/30
Inventor 陈秋云穆威宇肖新新
Owner 上海谱尼医学检验实验室有限公司
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