A nano-artificial antibody inhibitor for real-time imaging of tumor markers in living cells and its preparation method

An artificial antibody and real-time imaging technology, applied in the field of biomedicine, can solve the problems of unsatisfactory tumor suppression effect, unsatisfactory research and development effect, and high price, and achieve important application value, simplified operation, and high specificity

Active Publication Date: 2021-05-04
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At this stage, the research on this type of protein is gradually deepening, and inhibitors targeting these proteins have been developed to regulate the growth of tumors, but the effect of inhibiting tumors is not satisfactory
In addition, there are still problems such as low sensitivity and high price in the detection of tumor markers, and the unsatisfactory research and development effect of inhibitors of tumor regulation-related proteins

Method used

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  • A nano-artificial antibody inhibitor for real-time imaging of tumor markers in living cells and its preparation method
  • A nano-artificial antibody inhibitor for real-time imaging of tumor markers in living cells and its preparation method
  • A nano-artificial antibody inhibitor for real-time imaging of tumor markers in living cells and its preparation method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: prepare the nanoartificial antibody inhibitor of transferrin (TRF)

[0045] Preparation of CdTe@SiO 2 : First prepare NaHTe solution, mix 50.8mg (0.4mmol) tellurium powder and 37.8mg (1mmol) NaBH 4 Put it into a 25mL double-necked flask, evacuate it under nitrogen protection, and then add 10.0mL distilled water. The reaction mixture was reacted at 80° C. for 30 minutes under nitrogen to obtain a deep red clear NaHTe solution, which was used for subsequent reactions. 0.2mmol of CdCl 2 Add 0.34mmol of mercaptopropionic acid to 40mL of water and mix, add 1.0M NaOH solution drop by drop under stirring to adjust the pH to 11.9, vacuumize and fill with nitrogen, stir at room temperature, and inject 2.3mL of newly prepared NaHTe solution through a syringe added to the reaction system. The reaction mixture was then heated to reflux (100°C) to produce CdTe quantum dots, as figure 1 shown. Then take 30μl+470μL lye (0.45%wt ammonia water+15.9×10 -3 M NaOH), it...

Embodiment 2

[0047] Embodiment 2: the nano artificial antibody inhibitor of preparation human serum albumin (HSA)

[0048] First prepare the nano-gold cage: take 6mL of ethylene glycol in a 20mL glass bottle, put it in an oil bath, and heat it for 1h. Then add 80 μL of 3 mM Na to the reaction vial 2 S solution, after heating for 10min, add 1.5mL 1mg / mL PVP solution into the bottle, then immediately add 0.5mL 3mmol / L silver nitrate solution, and heat for half an hour. The reaction solution produced a lot of solids, the heating was stopped, and the reaction was taken out of the oil bath and cooled to room temperature. Transfer the reaction solution to a 50 mL centrifuge tube, rinse the vial with acetone twice the reaction volume, and transfer the wash solution to the centrifuge tube. Centrifuge, remove the supernatant, and wash three times with deionized water. Silver nanocubes can be obtained. Get 5mL 1mg / mL PVP aqueous solution in the round bottom flask of 50mL, add 100 μ L of above-me...

Embodiment 3

[0053] Embodiment 3: prepare the nano-artificial antibody inhibitor of horseradish peroxidase (HRP)

[0054] Add 50mg of chloroauric acid and 500mL of ultrapure water to a 2000mL Erlenmeyer flask, heat to 98°C under magnetic stirring, then add a certain amount of sodium citrate (Table 1), and maintain the temperature at 98°C for reaction. The color of the solution turned blue and then black within a few seconds, and then turned red again. After continuing to heat for 10 minutes, stop heating and continue stirring for 0.5 hours to prepare a nano-gold colloidal solution. After cooling at room temperature, high-speed centrifugation and purification, save for future use. The particle size of the synthesized gold nanoparticles can be changed by changing the amount of sodium citrate. The particle size and morphology of the synthesized nano-gold colloids were studied by transmission electron microscopy (TEM), such as Figure 8 shown.

[0055] Table 1

[0056]

[0057] A tempera...

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Abstract

The invention discloses a nanometer artificial antibody inhibitor for real-time imaging of tumor markers in living cells and a preparation method thereof. The nano-artificial antibody inhibitor is a molecularly imprinted polymer artificial antibody with biomimetic molecular recognition performance for tumor markers constructed on the surface of nanomaterials, which has the advantages of high specificity, good reproducibility, and good physical and chemical stability. Temperature or pH environmental response performance. Using the surface plasmon resonance effect of nano-gold or the fluorescence effect of quantum dots, the Raman spectrum quantitative detection or fluorescence quantitative detection of tumor markers can be realized, which has the advantages of fast, high sensitivity, label-free, non-destructive, and real-time quantitative detection. After nano-artificial antibody inhibitors are introduced into cells, real-time visualization and imaging of tumor markers can be realized in living cells, and through the selective capture and enrichment of proteins related to regulating tumor growth, they can inhibit tumor growth and can be used for early diagnosis and treatment of cancer. Targeted therapy.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a nano-artificial antibody inhibitor used for real-time imaging of tumor markers in living cells and a preparation method thereof. The obtained nano-artificial antibody inhibitor can realize the detection of tumor markers. Raman spectroscopy or fluorescence quantitative detection and real-time visualization imaging, and by inhibiting tumor growth. Background technique [0002] Tumor markers, also known as tumor markers, refer to substances that are characteristically present in malignant tumor cells, or abnormally produced by malignant tumor cells, or produced by the host in response to tumor stimuli, and can reflect the occurrence and development of tumors. A class of substances that monitor tumor response to therapy. Tumor markers exist in the tissues, body fluids and excreta of tumor patients, and can be detected by immunological, biological and chemical metho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574C08G73/06
CPCC08G73/0672G01N33/57496G01N2333/4704
Inventor 吕永琴张童李燕陈自慢谭天伟
Owner BEIJING UNIV OF CHEM TECH
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