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Magnetic nano material and preparation and application thereof

A magnetic nanometer and magnetic material technology, which is applied in the preparation method of peptides, the preparation of test samples, and other chemical processes to achieve the effect of good hydrophilicity, easy separation and high hydrophilicity

Inactive Publication Date: 2016-07-20
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a large number of hydrophilic materials have been used for the enrichment of glycopeptides, the enrichment sensitivity and specificity of hydrophilic materials still need to be improved

Method used

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  • Magnetic nano material and preparation and application thereof
  • Magnetic nano material and preparation and application thereof
  • Magnetic nano material and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1) Magnetic nanoparticles (Fe 3 o 4 SiO 2 -NH 2 )Synthesis.

[0034] 2.70g ferric chloride (FeCl 3 ·6H 2 O) be dissolved in the ethylene glycol of 100mL, mechanically stir to obtain yellow transparent liquid, then add 7.20g anhydrous sodium acetate (CH 3 COONa), after continuing to stir for 30 minutes, the solution of gained was transferred in the stainless steel reaction kettle of the Teflon liner of 200mL. After the reaction kettle was placed at 200°C for 8 h, the obtained material was washed three times with 30 mL of ethanol and 30 mL of water, and then vacuum-dried at 60°C to obtain Fe 3 o 4 (175nm).

[0035] Weigh 0.2gFe 3 o 4 Particles were dispersed into 50mL of dilute hydrochloric acid with a concentration of 0.1M, and ultrasonically treated for 25min. Then wash with deionized water to neutral Fe 3 o 4 The particles were dispersed into a mixture of 160mL ethanol, 40mL water and 2.0mL concentrated ammonia water, mechanically stirred for 25min, and 1....

Embodiment 2

[0040] Example 2 Analysis of Standard Glycoprotein Samples Based on Magnetic Nanomaterials Pretreated by Glycosylated Protein Samples

[0041] Preparation of glycoprotein samples: Add 1mg of human immunoglobulin G (IgG) to 1mL of 0.1M ammonium bicarbonate / 8M urea solution (pH8.2), 20 μL of 1M dithiothreitol (DTT), and mix well at 60 ℃ reaction 1h. Add 7.2 mg of iodoacetamide (IAA) to the above solution, and react for 40 min at room temperature under the condition of avoiding light. After adding 40 μL (40 μg) of trypsin solution, the system was reacted at 37° C. for 16 h. The resulting solution was desalted through a SPE column. The obtained protein solution was stored in a -30°C refrigerator for future use.

[0042] Glycosylated protein pretreatment and MALDI-TOFMS analysis based on magnetic nanomaterials: the process starts with the enrichment of glycosylated polypeptides. Evenly disperse 20 μg of the magnetic nanomaterial prepared in Example 1 into 400 μL of 88% ACN / 5% T...

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Abstract

The invention relates to a pretreatment process of a glycosylated protein sample, in particular to a novel magnetic nano material for pretreatment of the glycosylated protein sample.The method comprises the steps of preparation of the magnetic nano material, subsequent modification of glycopeptide dendritic macromolecules, and enrichment of glycosylated polypeptide with the hydrophilic interaction chromatography.The method is high in sensitivity and suitable for pretreatment of glycosylated protein in a biological sample.

Description

technical field [0001] The present invention relates to the synthesis of novel magnetic nanomaterials and the sample pretreatment of glycosylated proteins, in particular to a magnetic nanomaterial used for fast sample pretreatment of glycosylated proteins, which can be used for glycosylated polypeptides Enrichment. Background technique [0002] Protein post-translational modification (PTMs) is a hotspot in proteomics research in recent years. Protein glycosylation modification is the most common and one of the most important protein post-translational modifications. It plays a key regulatory role in many important life activities, such as protein folding, cell-cell interaction, molecular recognition, immune response and inflammatory response. In addition, abnormal protein glycosylation modification is also an important sign of the development of cancer and some other diseases. The diagnostic markers and therapeutic targets of many diseases come from glycosylated proteins....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/286B01J20/30C07K1/22G01N1/28
Inventor 邹汉法李吉楠王方军
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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