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Surface finish C8 alkyl chain magnetic silicon ball and preparing method and application thereof

A magnetic silica sphere and surface modification technology, which is applied to the preparation method of peptides, the preparation of test samples, chemical instruments and methods, etc., can solve the problems of cumbersome operation and achieve good practical value and application prospect

Inactive Publication Date: 2008-05-28
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Professor Yang Pengyuan's group successfully applied zeolite nanoparticles to the enrichment of trace peptides[5-7], but this method requires high-speed centrifugation to separate the sample and zeolite mixture, and the operation is relatively cumbersome

Method used

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  • Surface finish C8 alkyl chain magnetic silicon ball and preparing method and application thereof
  • Surface finish C8 alkyl chain magnetic silicon ball and preparing method and application thereof
  • Surface finish C8 alkyl chain magnetic silicon ball and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Synthesis of magnetic silicon spheres with surface-modified C8 alkyl chains

[0028] The synthesis of magnetic silicon sphere materials with surface-modified C8 alkyl chains is divided into three steps.

[0029] First, the aminoferroferric oxide magnetic nanoparticles were synthesized by hydrothermal method: 1.0 g FeCl 3 ·6H 2 O was dissolved in 30 mL of ethylene glycol, and magnetically stirred for 0.5 h to obtain a yellow transparent solution. Then 4.0 g of anhydrous NaAc was added, and after magnetic stirring for 0.5 h, a brown-yellow transparent solution was obtained. The resulting solution was transferred into a 200 mL Teflon-lined stainless steel reaction kettle. Put it in an oven at 200°C for 12 hours. After cooling to room temperature, the product was repeatedly washed with deionized water five times to remove water-soluble impurities such as sodium acetate and ethylene glycol, and finally the product was vacuum-dried at 50°C for future use.

[00...

Embodiment 2

[0032] Example 2 The use of magnetic silica spheres with surface-modified C8 alkyl chains for peptide enrichment

[0033] (1) Enzymatic hydrolysis of protein solution:

[0034] Take 1.0 mg protein sample, including bovine serum albumin (BSA), cytochrome C (Cytochrome C), horse myocardium (Myoglobin), egg albumin (Ovalbumin), bovine β-casein (β-casein) and milk The extracted casein (Casein) was dissolved in 1.0mL water respectively, and after denatured by heating, ammonium bicarbonate solution was added to the solution to adjust the pH of the system to about 8, and 25 μg trypsin. At a temperature of 37°C, the enzymatic hydrolysis was terminated after 12 hours, and the enzymatic hydrolysis solution was frozen in a -80°C refrigerator until use.

[0035] (2) Separation and enrichment of peptides:

[0036] Make 20 μL to a concentration of 2 mg mL -1 The magnetic material dispersion solution was added to 1mL with a concentration of 5fmol μL -1 In the standard peptide or the try...

Embodiment 3

[0041] Example 3 Surface modification of Fe with C8 alkyl chain 3 o 4 @SiO 2 Magnetic materials for the enrichment of peptides in human serum

[0042] Take 10 μL of adult serum, add 20 μL of PBS buffer solution to dilute, then add 2 μL of 10 mg / mL surface-modified C8 alkyl chain magnetic material; pipette 5 times, after 30 seconds, remove the supernatant by magnetic separation. The material enriched in peptides was washed three times with PBS buffer solution with a pH of 7.0, and the supernatant was removed; 5 μL of 50% (v / v) acetonitrile aqueous solution was added to elute the peptides enriched in the material, and magnetic separation was obtained. Eluate: Spot 0.5 μL of the eluate containing the enriched peptides onto the target plate, dry and then spot 0.5 μL of CHCA (5 mg / mL; dissolved in 50% (v / v) acetonitrile and 0.1% (v / v) in TFA). After drying, MALDI-TOF MS analysis was performed. The results of mass spectrometry identification are shown in Figure 10.

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Abstract

The invention belongs to the technical field of inorganic materials and analysis, in particular to a Fe3O4 at SiO2 magnetic silicon ball with the surface being decorated by C8 alkyl chain and the preparation method and application thereof. The magnetic nanometer silicon ball is acquired by synthesizing into the magnetic nanometer material of ferroferric oxide at first, and then synthesizing into the Fe3O4 at SiO2 magnetic micro-ball material with shell structure, and further adopting C8 to conduct surface chemical decorating. The magnetic nanometer particles of the fixing C8 are taken as micro-absorbent, the specific surface is large, the peptide segments in organism are enriched, and the method is simple and effective. The material has good utility value and potential application in proteomics and other fields.

Description

technical field [0001] The invention belongs to the technical field of inorganic materials and analysis, and in particular relates to a magnetic silicon sphere with a surface-modified C8 alkyl chain and a preparation method and application thereof. [0002] The magnetic silica sphere proposed by the present invention can be used as an adsorbent for separation and enrichment of trace polypeptides and direct analysis by MALDI-TOF / MS to enrich peptides in organisms. The material has good practical value and application prospect in proteomics and other fields. Background technique [0003] It is well known that protein composition is diverse and variable, that is, in different cells of the same species or in the same cell at different times, its protein composition is constantly changing, resulting in extremely complex protein composition. At the same time, due to the post-translational modification of proteins, one mRNA often corresponds to multiple proteins, that is, the numb...

Claims

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Application Information

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IPC IPC(8): B01J20/06B01J20/26C07K1/14G01N33/48G01N1/28
Inventor 邓春晖林华清戚大伟刘扬张祥民
Owner FUDAN UNIV
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