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A nano-biosensor using fluorescence detection and its preparation method and application

A biosensor and fluorescence detection technology, applied in the direction of fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of large differences in chemical composition, low sensing signal intensity, and limited source of components, achieving good uniformity and wide application Foreground, the effect of simplicity in the preparation process

Active Publication Date: 2021-03-26
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The present invention aims at the deficiencies of the prior art, and provides a nano-biosensor for detecting lactate dehydrogenase activity using fluorescence, a preparation method and its application, so as to overcome the limited sources, poor stability, and chemical composition of existing biosensor components. Due to the large difference and low sensing signal intensity, its application in the biological field is expanded; at the same time, the preparation process is simplified, the preparation cost is reduced, and it is easy to industrialize; because it overcomes the technical defects of existing products, it can realize the use of fluorescent signals to measure The detection and sensing of lactate dehydrogenase activity, the obtained sensor and its preparation process can meet the needs of cancer cell diagnosis and anticancer drug screening

Method used

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  • A nano-biosensor using fluorescence detection and its preparation method and application
  • A nano-biosensor using fluorescence detection and its preparation method and application
  • A nano-biosensor using fluorescence detection and its preparation method and application

Examples

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Embodiment 1

[0032] The nano-biosensor that uses fluorescence to detect the activity of lactate dehydrogenase provided in this example is a doped Mn with red light emission synthesized by a hydrothermal method. 2+ Based on the cubic phase rare earth up-conversion luminescent nanoparticles, it is made through the hydrophilic-hydrophobic interaction between the surface oleic acid ligand and the Nile blue derivative (expressed as NB-C18PMH-PEG), which is water-soluble. A hybrid nanobiosensor with stable structure, small difference in chemical composition, and high sensing signal intensity.

[0033] The preparation method of the above-mentioned nano-biosensor using fluorescence detection of lactate dehydrogenase activity, doped Mn with red light emission 2+ Rare earth up-conversion luminescent nanoparticles NaYbF 4 : Mn / Er as the basis, through the hydrophilic and hydrophobic interaction between its surface oleic acid ligand and Nile blue derivatives, its surface is functionally modified, spe...

Embodiment 2

[0048] A nano-biosensor that uses fluorescence to detect lactate dehydrogenase activity and its preparation method provided in this example is basically the same as Example 1, except that the doped Mn that emits red light 2+ Rare earth up-conversion luminescent nanoparticles NaYbF 4 : Mn / Er / Tm as the basis, including the following steps:

[0049] (1) Prepare 10 mL of oil-soluble doped Mn with oleic acid as the surface ligand 2+ Rare earth up-conversion luminescent nanoparticles NaYbF 4 : Mn / Er / Tm cyclohexane dispersion, prepare 10-20mL of absolute ethanol, mix at a volume ratio of 1:1-1:2 and ultrasonically disperse at room temperature, centrifuge at 8000-10000 revolutions per minute After washing, the resulting solid was dispersed in 10 mL of chloroform to obtain a first dispersion.

[0050] (2) First, ultrasonically dissolve 0.1-0.4g polymaleic anhydride / 1-octadecene alternating copolymer, 1.43-5.72g methoxypolyethylene glycol amine and 0.11-0.44g EDC in 100-250mL dichlor...

Embodiment 3

[0054] A nano-biosensor that uses fluorescence to detect lactate dehydrogenase activity and its preparation method provided in this example is basically the same as in Examples 1 and 2, except that the doped Mn that emits red light 2+ Rare earth up-conversion luminescent nanoparticles NaYF 4 : Yb / Mn / Er as the basis, including the following steps:

[0055] (1) Prepare 10 mL of oil-soluble doped Mn with oleic acid as the surface ligand 2+ Rare earth up-conversion luminescent nanoparticles NaYF 4 : Cyclohexane dispersion of Yb / Mn / Er, prepare 10-20mL of absolute ethanol, mix at a volume ratio of 1:1-1:2 and ultrasonically disperse at room temperature, centrifuge at 8000-10000 rpm After washing, the resulting solid was dispersed in 10 mL of chloroform to obtain a first dispersion.

[0056] (2) First, ultrasonically dissolve 0.1-0.4g polymaleic anhydride / 1-octadecene alternating copolymer, 1.43-5.72g methoxypolyethylene glycol amine and 0.11-0.44g EDC in 100-250mL dichloro into ...

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Abstract

The invention discloses a nano biosensor for detecting lactate dehydrogenase activity by fluorescence. The nano biosensor is characterized in that the nano biosensor is based on Mn2+-doped hydrothermally-synthesized cubic phase rare earth upconversion luminescent nano particles with red light emission, the nano biosensor is made through hydrophilic-hydrophobic interaction between the surface oleicacid ligand and Nile Blue derivatives (NB-C18PMH-PEG), and the nano biosensor has properties of water solubility, stable structure and small chemical composition difference and is a hybrid nano biosensor with high sensing signal intensity. The invention further provides a preparation method and an application thereof. The nano biosensor is advantaged in that good stability and good biological compatibility are achieved, and the nano biosensor is utilized for upconversion of fluorescence imaging signals and can play a role in assisting cancer cell diagnosis and drug screening in vitro and meets needs of clinical diagnosis and anticancer drug development. The preparation method has properties of concise preparation process, high efficiency and stable quality.

Description

technical field [0001] The invention relates to the technical field of manufacturing nano-biological materials, in particular to a nano-biosensor for detecting the activity of lactate dehydrogenase using fluorescence, a preparation method and an application thereof. Background technique [0002] For a long time, cancer is still one of the important causes of human death, and the research on cancer has a long history. German physiologist Otto Warburg proposed in 1924 that the occurrence of cancer is due to the enhanced anaerobic glycolysis of intracellular sugar combined with This reduction in oxygen consumption is known as the Warburg effect. Cancer cells typically fuel themselves by enhancing the anaerobic glycolysis of glucose, which is converted to lactate by lactate dehydrogenase. Therefore, the activity of lactate dehydrogenase in cancer cells is much higher than that in normal cells. And amide adenine dinucleotide (NAD+) is the coenzyme factor of lactate dehydrogenas...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 孙丽宁王卓姜宏施利毅张强
Owner SHANGHAI UNIV
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