Fluorescent glucose nano biosensor and preparation method thereof

A biosensor, glucose technology, applied in the field of biochemical analysis and quantitative research, to achieve the effect of high sensitivity, low toxicity and good stability

Inactive Publication Date: 2014-03-05
BEIJING JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it has not been reported that glucose oxidase molecules are coupled to the surface of oxygen nanosensors to construct glucose oxidase fluorescent nanosensors for rapid and accurate fluorescence detection of glucose in trace samples.

Method used

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  • Fluorescent glucose nano biosensor and preparation method thereof
  • Fluorescent glucose nano biosensor and preparation method thereof
  • Fluorescent glucose nano biosensor and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A fluorescent type glucose nanometer biosensor, which is composed of functionalized fluorescent nanoparticles, and has a structure such as figure 1 Shown. Firstly, a ratio fluorescent oxygen nanosensor coated with polylysine (PLL) shell layer was prepared by reprecipitation method: the inside is formed by hybridization of dodecyltrimethoxysilane (DTS) and polystyrene (PS) The core is doped with two fluorescent dye molecules (oxygen probe molecule platinum (Ⅱ) MESO-tetrakis (pentafluorobenzene) porphine (PtTFPP) and fluorescent reference molecule coumarin 6 (C6)); the exterior is PLL shell. The morphology and particle size of the oxygen nanosensor are as figure 2 As shown, the particle size is concentrated around 258.2nm. Then, based on the Schiff base reaction between primary amine and glutaraldehyde, glucose oxidase is coupled to the surface of the oxygen nanosensor, so that it can catalyze the oxidation of glucose. The specific coupling reaction principle is as follo...

Embodiment 2

[0036] PtTFPP, C6, PS and DTS were dissolved in tetrahydrofuran in a mass ratio of 2:2:46:50, and their total concentration in the solution was 0.2%. Then, use a micro-adjusting syringe to take 500 μL of the solution, and quickly inject it into deionized water with a PLL concentration of 0.02 mg / ml with a pH of 9 (adjust the pH with ammonia water) under ultrasonic vibration, and the resulting suspension is ultrasonically After shaking for 2 hours, followed by dialysis in deionized water for 24 hours, PLL-coated core-shell structured nanoparticles were obtained, which are sensitive to oxygen, and the fluorescence intensity decreases with increasing oxygen concentration. This is the fluorescent nano oxygen sensor.

[0037] In the fluorescent nano-oxygen sensor prepared above, that is, the PLL-coated nanoparticles, first add the same amount of glutaraldehyde as the PLL substance, shake for 5 minutes at 25°C, and mix it uniformly; then add glucose Oxidase (GO x ) In the aqueous solu...

Embodiment 3

[0043] 1) Dissolve platinum (II) MESO-tetrakis (pentafluorobenzene) porphine, coumarin 6, polystyrene and dodecyltrimethoxysilane with a mass ratio of 1:2:47:50 in tetrahydrofuran , The total concentration in the solution is 0.1%;

[0044] 2) Prepare a deionized aqueous solution of polylysine with a concentration of 0.02 mg / ml, and adjust the pH of the deionized aqueous solution of polylysine to 8 with ammonia;

[0045] 3) Take 300 μL of the tetrahydrofuran solution described in step 1) and inject it into the deionized aqueous solution of polylysine described in step 2) under ultrasonic vibration to obtain a suspension. After ultrasonic vibration for 1 hour, it is then deionized After 12 hours of dialysis in water, fluorescent oxygen nanoparticles with core-shell structure are obtained, and the particle size is about 240nm;

[0046] 4) In step 3) the fluorescent oxygen nanoparticles with core-shell structure, first add glutaraldehyde in the amount of polylysine and other substances,...

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Abstract

The invention discloses a fluorescent glucose nano biosensor. Active ingredients of the sensor are functionalized fluorescent nanoparticles. The fluorescent glucose nano biosensor is prepared by combining an enzyme catalysis method and a fluorescent oxygen nano sensor. Fluorescence detection of glucose is indirectly realized based on consumption of glucose oxidase on oxygen in a glucose catalysis process. The glucose nano biosensor has the advantages of high response speed, high sensitivity and high stability, and the lower limit of glucose detection can be 0.5mmol/L. Because the sensor has a nanoscale size and high biocompatibility, the sensor can be used for detecting the concentration of glucose in microenvironments (such as living cells) in organisms and in human body fluid (such as plasma).

Description

Technical field [0001] The invention belongs to the technical field of biochemical analysis and quantitative research, and specifically relates to a fluorescent glucose nano biosensor and a preparation method thereof. Background technique [0002] The measurement of glucose concentration has a wide range of applications in clinical analysis, biotechnology, agriculture, food industry, etc. Among them, in clinical analysis, it is very important for the diagnosis of diabetes, subsequent treatment and detection of cell metabolism. Diabetes is a very common disease. Insulin deficiency in patients leads to disorders of blood glucose metabolism. Therefore, the glucose concentration in the blood changes more intensely than in the normal body. A series of technologies for detecting glucose have been developed. [0003] Glucose detection methods mainly include spectrophotometry, optical rotation, gas chromatography, colorimetry and so on. Among them, the spectrophotometric method require...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/52
Inventor 彭洪尚高少伟
Owner BEIJING JIAOTONG UNIV
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