Method for synthesizing carbon quantum dot by using biologic active enzyme auxiliary microwave method, carbon quantum dot prepared by using method, 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 not having the function of cancer diagnosis and treatment, not being able to enter the nucleus, and low yield, so as to 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 RNase A as biosynthetic template, super-fluorescent carbon quantum dots were prepared.
[0041] Step 1: At room temperature, weigh 1 g of citric acid and 20 mg of RNase A, and measure 5 mL 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 RNase A, namely Super fluorescent carbon quantum dots modified by RNase A...
Embodiment 2
[0049] Using glucose as precursor and RNase A as biosynthetic template, high-intensity fluorescent carbon quantum dots were prepared.
[0050] Step 1: At room temperature, weigh 1g glucose and 10mg RNase A, measure 5mL 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 brown-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 RNase A. RNase A modified carbon quantum dots were obtained, and the yield of RN...
Embodiment 3
[0055] Laser confocal imaging of gastric cancer MGC-803 cells using RNase AC-dots as fluorescent labels.
[0056] Step 1: Prepare multifunctional fluorescent RNase AC-dots probe labeled with BRCAA1 antibody. Since the surface of RNase AC-dots is rich in carboxyl and amino groups at the same time, the -COOH on the surface of RNase AC-dots particles is activated with cross-linking agents carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The -NH2 on the surface of the BRCAA1 antibody and the -COOH on the surface of the quantum dots were condensed 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 logarithmic growth phase we...
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