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
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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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