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Porous carbon carrier, preparation method thereof and application of porous carbon carrier in ligand fishing

A technology of porous carbon and carrier, applied in the field of porous carbon carrier and its preparation, can solve the problems of limited loading rate, limited development and application of ligand fishing method, insufficient stability, etc., and achieve good stability, repeatability, and screening efficiency. High, specific effect

Pending Publication Date: 2022-03-25
MACAU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the micro-nano materials used to immobilize biomolecules in the existing ligand fishing methods still have defects such as limited loading rate and insufficient stability, which limit the development and application of ligand fishing methods to a certain extent.

Method used

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  • Porous carbon carrier, preparation method thereof and application of porous carbon carrier in ligand fishing
  • Porous carbon carrier, preparation method thereof and application of porous carbon carrier in ligand fishing
  • Porous carbon carrier, preparation method thereof and application of porous carbon carrier in ligand fishing

Examples

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Effect test

Embodiment 1

[0046] The present embodiment provides a porous carbon carrier, which is a hierarchical porous carbon with single-atom Zn sites, named SAZn-HPC, and its preparation method includes the following steps:

[0047] (1) ZnCl 2 Dissolve with sodium polyacrylate into deionized water under ultrasonic stirring to obtain a mixture;

[0048] (2) freeze-drying the mixture obtained in step (1), and then place it in Ar gas for carbonization to obtain initial product;

[0049] (3) grinding, washing and drying the initial product obtained in step (2) to obtain the porous carbon carrier SAZn-HPC.

[0050] figure 1 Shown is the morphological characteristic map of the above porous carbon support (SAZn-HPC), which is represented byfigure 1 It can be seen that a broad peak at 23° was observed in SAZn-HPC pointing to the (002) peak of amorphous carbon (JCPDS No. 41-1487), indicating the existence of a random combination of graphite and turbostratic stacking ( figure 1 A); from scanning electron ...

Embodiment 2

[0053] This example provides a ligand screening system, including β-amyloid (amyloidβ-protein, Aβ) and the porous carbon carrier SAZn-HPC prepared in Example 1, named Aβ@SAZn-HPC. The preparation method of the ligand screening system comprises the following steps:

[0054] (1) Dissolve Aβ in hexafluoroisopropanol for ultrasonic treatment for 15 min, then dry in a nitrogen blower to obtain an Aβ film, and store in a -80°C refrigerator for later use. The resulting A[beta] films were redissolved with DMSO and PBS, and the A[beta] was incubated at 37[deg.]C for 5 days to form the aggregated form of A[beta] for assay.

[0055] (2) Dilute Aβ with PBS to different concentrations (0.125mg / mL, 0.250mg / mL, 0.500mg / mL), then mix it with the SAZn-HPC prepared in Example 1, and place it in a shaker at 4°C overnight , washed three times with 4.0 mL PBS, and centrifuged at 1000 × g for 10 min to prepare Aβ@SAZn-HPC.

Embodiment 3

[0065] The present embodiment provides a method for screening Aβ potential inhibitors from turmeric extract, comprising the following steps:

[0066] (1) pulverize the traditional Chinese medicine turmeric, add 75% (v / v) ethanol for reflux extraction for 2 hours, the filtrate is evaporated to dryness under reduced pressure in a rotary evaporator, and stored at 4°C;

[0067] (2) The Aβ@SAZn-HPC prepared in Example 2 was vortexed and mixed with the turmeric extract in step (1), the supernatant was removed, incubated at 37° C. for 30 min, washed with PBS for 3 times and mixed with 70% Methanol was mixed by vortexing, and the obtained solution was subjected to mass spectrometry detection and analysis; meanwhile, PBS was used instead of turmeric extract for control experiments.

[0068] Conditional parameters for mass spectrometry detection include:

[0069] Mobile phase composition:

[0070] Mobile phase A: 0.1% (v / v) aqueous formic acid; Mobile phase B: acetonitrile;

[0071] ...

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Abstract

The invention belongs to the technical field of ligand fishing, and discloses a porous carbon carrier, a preparation method thereof and application of the porous carbon carrier in ligand fishing. The preparation method of the porous carbon carrier comprises the following steps: (1) dissolving Zn salt and polyacrylate in water to obtain a mixture; and (2) freeze-drying the mixture, and carbonizing the freeze-dried mixture in inert gas to obtain the porous carbon carrier. The porous carbon carrier has good capability of loading and fixing protein, can be used for a ligand fishing technology, shows good stability and repeatability, and can be used for rapidly, efficiently and accurately screening target active substances from natural plant components.

Description

technical field [0001] The invention belongs to the technical field of ligand fishing, and particularly relates to a porous carbon carrier, a preparation method thereof, and an application in ligand fishing. Background technique [0002] Ligand fishing is based on intermolecular affinity to identify interacting ligands and receptors. By combining with modern organic analysis methods, it opens up a new way to study the interaction between biological macromolecules and ligands. . In short, ligand fishing is based on the interaction between drug targets and active ligands, and the active ligands are "fished" out of complex sample systems. The ligand-fishing strategy with high sensitivity and high-throughput assay has broad prospects in the screening of active compounds. The current research results show that the ligand-fishing technology can realize the rapid screening of active products from complex extract components. Preferred strategy for the directed isolation of native ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/88G01N1/28
CPCG01N30/06G01N30/88G01N1/28
Inventor 张伟姜志宏周明月吴璇王彩云
Owner MACAU UNIV OF SCI & TECH
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