A method for synthesizing carbon quantum dots by biologically active enzyme-assisted microwave method, carbon quantum dots prepared therefrom and application thereof

A technology of biologically active enzymes and carbon quantum dots, which is applied in the fields of nanomaterial science and biomedical engineering, can solve the problems of no cancer diagnosis and treatment function, inability to enter the nucleus, and low yield, and achieve low synthesis cost, high yield, The effect of simple equipment

Inactive Publication Date: 2016-06-01
SHANGHAI JIAOTONG UNIV
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  • Abstract
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Problems solved by technology

[0008] In view of the low yield of carbon quantum dots in the prior art and the prepared carbon quantum dots, if no further modification of the target molecule, basically cannot enter the nucleus, and do not have the function of diagnosis and treatment of cancer and other defects, the present invention provides a Method for preparing carbon quantum dots with high yield, good biological safety, simple synthesis method and low cost

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  • A method for synthesizing carbon quantum dots by biologically active enzyme-assisted microwave method, carbon quantum dots prepared therefrom and application thereof
  • A method for synthesizing carbon quantum dots by biologically active enzyme-assisted microwave method, carbon quantum dots prepared therefrom and application thereof
  • A method for synthesizing carbon quantum dots by biologically active enzyme-assisted microwave method, carbon quantum dots prepared therefrom and application thereof

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Embodiment 1

[0040] Using citric acid as carbon precursor and RNaseA as biosynthetic template, super-fluorescent carbon quantum dots were prepared.

[0041] Step 1: At room temperature, weigh 1g of citric acid and 20mg of RNaseA, and measure 5mL of H 2 0, all moved into a 25mL glass reaction bottle, and the reaction bottle was placed in an ultrasonic wave with a working frequency of 40kHz and a power of 200W for 1min to obtain a uniform aqueous solution;

[0042] Step 2: Put the aqueous solution obtained in Step 1 in a household microwave oven, and microwave it for 4 minutes at a power of 800W, and obtain a brown-yellow liquid after reaction;

[0043] Step 3: Use a dialysis bag with a molecular weight cut-off of 5000 to dialyze the brownish-yellow liquid obtained in Step 2. The dialysis time is 24 hours. During the dialysis process, the ultrapure water is continuously replaced to remove unreacted citric acid and RNaseA. RNaseA-modified super-fluorescent carbon quantum dots were obtained, ...

Embodiment 2

[0049] Using glucose as precursor and RNaseA as biosynthetic template, high-intensity fluorescent carbon quantum dots were prepared.

[0050] Step 1: At room temperature, weigh 1g of glucose and 10mg of RNaseA, and measure 5mL of H 2 O, all were moved into a 25mL glass reaction bottle, and the reaction bottle was placed in an ultrasonic wave with a working frequency of 40kHz and a power of 200W for 1min to obtain a uniform aqueous solution.

[0051] Step 2: Put the aqueous solution obtained in Step 1 in a household microwave oven, and heat it in a microwave for 4 minutes at a power of 800W. After the reaction, a brown-yellow liquid is obtained.

[0052] Step 3: Use a dialysis bag with a molecular weight cut-off of 5000 to dialyze the brownish-yellow liquid obtained in Step 2. The dialysis time is 36 hours. During the dialysis process, the ultrapure water is continuously replaced to remove unreacted glucose and RNaseA, and you can get RNaseA modified carbon quantum dots, RNase...

Embodiment 3

[0055] Laser confocal imaging of gastric cancer MGC-803 cells using RNaseAC-dots as fluorescent labels.

[0056] Step 1: Prepare multifunctional fluorescent RNaseAC-dots probe labeled with BRCAA1 antibody. Since the surface of RNaseAC-dots is rich in carboxyl and amino groups at the same time, the -COOH on the surface of RNaseAC-dots particles was activated with cross-linking agents carbodiimide (EDC) and N-hydroxysuccinimide (NHS), through BRCAA1 antibody The condensation reaction of -NH2 on the surface and -COOH on the surface of the quantum dots was carried out for 2 hours to realize the coupling of the BRCAA1 antibody to the quantum dots. After three washes with PBS buffer, the obtained BRCAA1 antibody-labeled quantum dots were stored at 4°C for future use.

[0057] Step 2: Targeted enrichment of tumor cells. Gastric cancer MGC-803 cells were cultured by conventional in vitro culture methods, and the specific method was as follows: Gastric cancer MGC-803 cells in logarit...

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Abstract

The invention discloses a method for synthesizing a carbon quantum dot by using a biologic active enzyme auxiliary microwave method. The method specifically comprises the following steps: (1) ultrasonically dispersing a carbon precursor and biologic active enzyme into deionized water, so as to prepare a transparent solution; (2) putting the solution obtained in the step (1) into a domestic micro-oven for heating through microwave, and reacting to obtain claybank liquid which is the biologic active enzyme modified carbon quantum dot; (3) dialyzing the carbon quantum dot solution obtained from the reaction in the step (2) by using a dialysis bag, so as to remove unreacted carbon precursor and biologic active enzyme. The method is high in yield; as the prepared carbon quantum dot is modified by the biologic active enzyme, the carbon quantum dot can enter cell nucleus and has the diagnosis and treatment function on cancer; the biologic active enzyme is used as an active agent and a passivating agent in the reaction, and citric acid, citrate, glucose, fructose or amylase is taken as a carbon precursor, so that the fluorescence carbon quantum dot has good biologic security in the preparation and application process; and the cost is low since the microwave synthesis method is used.

Description

technical field [0001] The invention belongs to the fields of nanometer material science and biomedical engineering, and in particular relates to a method for synthesizing fluorescent carbon quantum dots by a biologically active enzyme-assisted microwave method. Background technique [0002] With the rapid development of nanotechnology and optical imaging technology, the use of unique optical, magnetic and electrical properties of nanomaterials has brought new opportunities for early diagnosis and treatment of major diseases in clinical medicine. [0003] Molecular imaging is one of the important means of clinical tumor detection. The development of biological nanoprobe technology provides new ideas for the development of molecular imaging technology. For example, AlivisatosP. published a paper entitled Nanocrystals in the journal Nature Biotechnology (2004, 22 (1): 47). The use of nanocrystals in biological detection (The use of nanocrystals in biological detection), this ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C01B31/02C09K11/65G01N21/64B82Y20/00C01B32/15
Inventor 沈广霞刘慧旸王青崔大祥
Owner SHANGHAI JIAOTONG UNIV
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