A kind of hydrophilic branch and c18 modified magnetic graphene material and its preparation method and application

A magnetic graphene and graphene technology, applied in the biological field, can solve problems such as low detection limit

Active Publication Date: 2020-11-20
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has successfully improved the enrichment efficiency of proteins, and has strong universality for proteins, and also has a very low detection limit.

Method used

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  • A kind of hydrophilic branch and c18 modified magnetic graphene material and its preparation method and application
  • A kind of hydrophilic branch and c18 modified magnetic graphene material and its preparation method and application
  • A kind of hydrophilic branch and c18 modified magnetic graphene material and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: magG@SiO 2 @PAMAM / C18 material synthesis

[0045] (1) Synthesis of magnetic graphene (magG): disperse 400-600 mg of graphene into 50-70 mL of concentrated nitric acid, reflux in a water bath at 50-70 °C for 6-10 hours; After washing with water until the solution is neutral, vacuum-dry it at 40-60°C, place it for storage, and obtain acidified graphene; then add 400-500 mg FeCl 3 • Dissolve 6H2O in 40-60 mL of ethylene glycol, stir to obtain a yellow transparent solution; dissolve the acidified graphene in the previous step in the obtained yellow solution, ultrasonic 1.0-2.0 The ratio is: 150-200 mg: 40-60 mL ; Then under the condition of ultrasonic stirring, add trisodium citrate, sodium acetate, polyethylene glycol (20000) and stir thoroughly for 1.0-1.5 hours, among them, trisodium citrate, sodium acetate, polyethylene glycol (20000) The ratio is: (150-180 mg): (1.5-2.0 g): (1.0-1.5 g); transfer the mixed solution to a hydrothermal reactor, and heat at 180...

Embodiment 2

[0049] Example 2: 100-200 µg magG@SiO2@PAMAM / C18 material is used for protein enrichment

[0050] Take 100-200 µg magG@SiO2@PAMAM / C18 and wash 3-4 times with ultrapure water. Add ultrapure water and an appropriate amount of protein mixture to make the total volume 95-110 µL; enrich at 20-30°C for 20 min-30 min; remove the supernatant by magnetic separation, and then wash with ultrapure water 2-3 times, Finally, use the eluent (80%-85% acetonitrile and 0.1-0.2% TFA solution) to elute for 20-25 min at 30-37°C to elute the enriched protein. The eluate is subjected to HPLC and MALDI-TOF MS analysis and detection.

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Abstract

The invention belongs to the technical field of biotechnology and specifically relates to a hydrophilic branch and C18 long-chain modified magnetic graphene material as well as a preparation method and application thereof in protein enrichment. The method comprises the following steps: utilizing a hydrothermal method to compound a high-stability magnetic graphene material; coating a layer of silicon dioxide on the surface according to a reversed-phase micro-emulsion method; adding amino and C18 silylation reagent; and connecting PAMAM with amino according to a glutaraldehyde crosslinking method, thereby acquiring the hydrophilic-hydrophobic dendritic molecule and C18 long-chain modified magnetic graphene material. The modified PAMAM molecule has higher hydrophilicity, so that the dispersibility of the material in water is better; the C18 long chain has higher hydrophobicity and is easy to enrich protein; in the enrichment for four standard proteins, the recovery rate is as high as above 80%; in the enrichment for standard protein BSA and myohemoglobin MYO, the limit of detection is as low as 1 ng / muL; the hydrophilic branch and C18 modified magnetic graphene material is practical and efficient, is high in repeatability, is high in stability and is wide in application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a hydrophilic dendritic molecule and a C18 long-chain modified magnetic graphene material, a preparation method thereof and an application in protein enrichment. Background technique [0002] With the completion of the genome sequencing project, life science has achieved great progress, and at the same time entered the post-genome era, and proteomics has also become a research frontier and hot spot in the post-genome era. Proteins are the executors of life functions and the direct manifestations of life phenomena. The study of protein structure and function will directly explain the mechanism of changes in life under physiological or pathological conditions. In recent years, the separation and identification of proteins has become a very important part of proteomics. A series of research methods based on liquid chromatography and biological mass spectrometry have been gr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/26B01J20/28B01J20/30C07K1/14
CPCB01J20/103B01J20/20B01J20/26B01J20/28009C07K1/14
Inventor 高明霞王亚楠张祥民
Owner FUDAN UNIV
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