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Preparation of organic phosphonic acid functionalized magnetic nano material and application in protein enrichment

An organic phosphonic acid and magnetic nanotechnology, which is applied in the fields of organic material/organic magnetic material magnetism, material separation, material analysis, etc., can solve the problems of cumbersome steps and non-green, and achieve the ability to bind and enrich proteins. The effect is good, the synthesis method is simple

Inactive Publication Date: 2013-03-27
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation of the above materials either requires high temperature, or the steps are cumbersome and non-green

Method used

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  • Preparation of organic phosphonic acid functionalized magnetic nano material and application in protein enrichment
  • Preparation of organic phosphonic acid functionalized magnetic nano material and application in protein enrichment
  • Preparation of organic phosphonic acid functionalized magnetic nano material and application in protein enrichment

Examples

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Effect test

Embodiment 1

[0040] Embodiment 1, Zr 4+ -Synthesis of NO2PA-MS and enrichment and detection of phosphorylated proteins Weigh 2 mg of magnetic nanoparticles coated with aminosilane, disperse them in 1 mL of secondary water under the action of ultrasound, and then weigh 6 mg of NO2PA, EDC and NHS , dissolved in 1 mL of PBS buffer solution (pH 5.6), added and mixed, then ultrasonically reacted at room temperature for 30 minutes. Then, under an external magnetic field, wash and separate, and wash three times with 1 mL of secondary water. Then add 1 mL of 50 mM ZrOCl 2 Hydrochloric acid solution (0.2M), stirred vigorously at room temperature for 20 minutes. Then wash three times with 0.2M hydrochloric acid solution and secondary water respectively.

[0041] Aspirate 50μL (1mg / mL) of the prepared Zr 4+ -NO2PA-MS particle suspension, add 50μL (10 -7 M or 10 -8M) β-casein digestion solution, shake and mix at room temperature for 30 seconds. Then, under an external magnetic field, fully wash...

Embodiment 2

[0043] Example 2, Zr 4+ -Synthesis of NC2P-MS and enrichment and detection of phosphorylated proteins

[0044] Weigh 1 mg of magnetic nanoparticles wrapped with aminosilane, disperse them in 1 mL of secondary water under ultrasonication, then weigh 3 mg of NC2P and dissolve them in 1 mL of LPBS buffer solution (pH 5.6), weigh 3 mg of EDC and 3 mg of NHS, and add to the mixture Afterwards, ultrasonic reaction was carried out at room temperature for 30 minutes. Then, under an external magnetic field, wash three times with 1 mL of secondary water. Then add 500 μL of 50 mM ZrOCl 2 Hydrochloric acid solution (0.2M), stirred vigorously at room temperature for 20 minutes. Then wash three times with 0.2M hydrochloric acid solution and secondary water respectively.

[0045] Draw 50μL (1mg / mL) of the prepared Zr 4+ -NC2P-MS suspension, add 50μL (10 -7 M or 10 -8 M) β-casein digestion solution, shake and mix at room temperature for 30 seconds. Then, under an external magnetic fie...

Embodiment 3

[0047] Example 3, Zr 4+ -Synthesis of CP-MS and its application in the enrichment and detection of phosphorylated proteins

[0048] Weigh 1mg of magnetic nanoparticles wrapped with aminosilane, disperse in 1mL of secondary water under the action of ultrasound, then add PBS buffer solution (pH 5.6) containing 3mg of CP, then weigh 3mg of EDC and 3mg of NHS, add the above mixture, Sonicate for 30 minutes at room temperature. Then, under an external magnetic field, wash three times with 1 mL of secondary water. Then add 500 μL of 50 mM ZrOCl 2 Hydrochloric acid solution (0.2M), stirred vigorously at room temperature for 20 minutes. Then wash three times with 0.2M hydrochloric acid solution and secondary water respectively.

[0049] Draw 50μL (1mg / mL) of the prepared Zr 4+ -CP-MS suspension, add 50μL (10 -7 M or 10 -8 M) β-casein digestion solution, shake and mix at room temperature for 30 seconds. Then, under an external magnetic field, fully washed three times with 200 μ...

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Abstract

The invention provides an R-CH2-PO3-M modified magnetic nanoparticle as a phosphorylation protein enrichment material, as well as a preparation method and application thereof in phosphorylation protein enrichment. The purpose of combining the functions of magnetic nanoparticles and phosphonic acid groups can be achieved through modifying carboxyl groups for organic phosphonic acid gene R. The synthesis method of the magnetic nano material with functionalized organic phosphonic acid is simple, rapid and green. The material has favorable enrichment effect on phosphorylation proteins, and an MALDI-TOF MS (Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry) can be used for detecting a phosphorylation peptide segment in 10-8M of bovine serum beta-casein digestive juice. The results show that a type of favorable phosphorylation protein enrichment materials can be constructed through combining the magnetic nanoparticles with the organic phosphonic acid.

Description

technical field [0001] The invention relates to the field of magnetic nanometer materials, and is a novel functionalized magnetic nanometer material which integrates the functions of magnetic nanometer particles and organic phosphonic acid. Background technique [0002] Due to their unique physical and chemical properties, such as high specific surface area and easy separation, magnetic nanomaterials have been widely used in drug delivery, bioseparation and other fields. Organic phosphonic acid (R-CH 2 -PO 3 h 2 ) chemistry is characterized by the chemical modification of the R group to make it a functional molecule with a phosphonic acid group and other groups at the same time, achieving chemical regulation to meet different functions. [0003] The reversible phosphorylation process of protein is the most common and important form of protein post-translational modification, which is related to a variety of life activities. However, due to the low abundance of phosphoryl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): H01F1/42B22F1/02G01N30/06
Inventor 苏延辉郭琳郑丽敏
Owner NANJING UNIV
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