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Nano-liposome encapsulating graphene quantum dots, and preparation and application of nano-liposome in biological enzyme activity detection

A technology of graphene quantum dots and nanoliposomes, which is applied in measurement devices, fluorescence/phosphorescence, instruments, etc., can solve the problems of difficult removal of surfactants, adverse effects of biological substances, and poor stability of antigen and antibody, and achieve luminescence. Long-term stability, good biological application potential, and the effect of reducing detection costs

Active Publication Date: 2020-08-07
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the added surfactant for phospholipid vesicle rupture is not easy to remove from the reaction system, and its introduction not only brings impurities to the system and complicates the reaction process, but even has adverse effects on biological substances
Although the method of using liposomes for enzyme-linked immunoassay has the advantage of strong affinity, there are still defects such as poor stability of antigen and antibody and high cost.
These also limit the further development of liposomes in the field of biosensors

Method used

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  • Nano-liposome encapsulating graphene quantum dots, and preparation and application of nano-liposome in biological enzyme activity detection
  • Nano-liposome encapsulating graphene quantum dots, and preparation and application of nano-liposome in biological enzyme activity detection
  • Nano-liposome encapsulating graphene quantum dots, and preparation and application of nano-liposome in biological enzyme activity detection

Examples

Experimental program
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Effect test

Embodiment 1

[0069] This embodiment provides a method for synthesizing graphene quantum dots.

[0070] Weigh 0.4g carbot vulcan XC-72 carbon black (brand: Mecoxlane, purchased from Guangzhou Puzhi Biotechnology Co., Ltd.), add to 100mL 6mol / L HNO 3 , stirred and refluxed for 24 hours under the condition of 130° C. (oil bath). Then the reacted solution was cooled to room temperature, the supernatant was sucked, and the acid was removed by heating until the pH was 5-7, and the final solution volume was 50 mL, which was named solution 1. The obtained solution 1 was filtered twice with filter paper (medium-speed qualitative filter paper (speed 102), pore size: 30-50 microns, brand: Biorad, Beijing Benowei Biotechnology Co., Ltd.) to obtain solution 2. Solution 2 was filtered again with a needle filter of 0.22 μm to obtain solution 3 . Solution 3 was centrifuged at 8000 rpm for 10 minutes, and then the supernatant was pipetted into an ultrafiltration centrifuge tube (pore size 3000 Da) to obt...

Embodiment 2

[0074] This embodiment provides a method for synthesizing liposomes.

[0075] Lecithin and cholesterol were mixed at a ratio of 5:1 (molar ratio, a total of 43.2mmol, 30mg), dissolved in 4ml of chloroform, and ultrasonically (power 100W) for 5 minutes to disperse evenly. Subsequently, the organic solvent was removed by rotary evaporation at 40° C. under reduced pressure for 1 hour by a rotary evaporator, and a layer of transparent film was uniformly formed at the bottom of the flask. Now add the graphene quantum dot solution of 2mL 0.1mg / ml (the graphene quantum dot prepared by embodiment 1 is dissolved in the phosphate buffer solution (pH7.0)), ice-bath ultrasonic (power 100W) 50 minutes, obtain milky white Cloudy liquid. Pass it through a 200nm polycarbonate membrane (that is, the filter membrane used by the liposome extrusion instrument has a pore size of 200nm), and repeatedly extrude 21 times (40° C.). Finally, the obtained liposome solution is dialyzed with a dialysis ...

Embodiment 3

[0078] Liposomes synthesized with different proportions of lecithin and cholesterol

[0079]The synthesis method is the same as in Example 2, except that lecithin and cholesterol are mixed in a ratio of 1:1 (molar ratio, 52.37mmol in total, 30mg) or 5:2 (molar ratio, 45.99mmol in total, 30mg).

[0080] Its scanning electron microscope results and particle size distribution are as follows Image 6 with 7 shown.

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Abstract

The invention discloses a nano-liposome encapsulating graphene quantum dots, and preparation and application of the nano-liposome in biological enzyme activity detection. The method comprises the following steps: (1) adding lecithin and cholesterol into chloroform, carrying out ultrasonic treatment to uniformly disperse the lecithin and cholesterol, and carrying out rotary evaporation to remove the chloroform, so as to obtain a liposome film; (2) adding a graphene quantum dot solution into the liposome film, and carrying out uniform ice-bath ultrasonic dispersion to obtain a mixed solution I;repeatedly extruding the mixed solution through a polycarbonate film to obtain a mixed solution II; and dialyzing the mixed solution II to obtain the nano-liposome encapsulating the graphene quantum dots. The nano-liposome prepared by the method is high in biological safety and long-acting and stable in luminescence; the liposome is broken in the presence of phospholipase A2 to release fluorescence signal molecules wrapped in the liposome, so that the liposome can be used for detecting the activity of pancreatitis marker phospholipase A2, and has a wide application prospect in the aspects of biological detection, clinical diagnosis and the like.

Description

technical field [0001] The invention belongs to the field of detection of organic / inorganic composite optical probe materials and disease-related markers, and in particular relates to a nano-liposome encapsulating graphene quantum dots, its preparation and its application in the detection of biological enzyme activity. Background technique [0002] Phospholipase is widely distributed in the human body and plays an important role in physiological processes such as phospholipid renewal, cell membrane remodeling, exocytosis and information transmission. Phospholipases mainly include phospholipases A1, A2, B1, B2, C and D, which will specifically act on each ester bond inside the phospholipid molecule to form different products. Among them, phospholipase A2 (PhospholipaseA 2 , PLA 2 ) can hydrolyze phospholipid sn-2 ester bonds to produce free fatty acids and lysophospholipids, which play a key role in pathophysiological processes such as information transmission and membrane ...

Claims

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Application Information

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IPC IPC(8): C09K11/65C09K11/02G01N21/64
CPCC09K11/025C09K11/65G01N21/6428
Inventor 李楠査勇超张美莹薛巍崔鑫王宇周平
Owner JINAN UNIVERSITY
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