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Polydopamine modified magnetic ball, synthetic method of nanomaterial by fixing Ti<4+> on the surface and application thereof

A polydopamine and surface immobilization technology, applied in the direction of material analysis, analysis of materials, and inorganic material magnetism by electromagnetic means, can solve the problems of loss of low-abundance phosphorylated peptides, time-consuming and inconvenient, and achieve good biocompatibility, The effect of simple fixing method and simple synthesis method

Inactive Publication Date: 2013-06-12
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional metal ion adsorption chromatography often requires high-speed centrifugation during sample preparation, which is not only time-consuming and inconvenient, but also leads to the loss of low-abundance phosphorylated peptides

Method used

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  • Polydopamine modified magnetic ball, synthetic method of nanomaterial by fixing Ti&lt;4+&gt; on the surface and application thereof
  • Polydopamine modified magnetic ball, synthetic method of nanomaterial by fixing Ti&lt;4+&gt; on the surface and application thereof
  • Polydopamine modified magnetic ball, synthetic method of nanomaterial by fixing Ti&lt;4+&gt; on the surface and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Polydopamine modified magnetic spheres and immobilized Ti on the surface 4+ Synthesis of Nanomaterials:

[0031] (1) Disperse 10 mg of magnetic spheres into 40 mL of dopamine aqueous solution with a concentration of 1 mg / mL, and make it disperse evenly;

[0032] (2) Under the condition of continuous mechanical stirring, quickly add 10 mL concentration of tris buffered solution of 10 mmol / L (pH is 8.5) to the product obtained in step (1);

[0033] (3) The solution obtained in step (2) was mechanically stirred at room temperature for 10-16 hours;

[0034] (4) washing the product obtained in step (3) three times with deionized water to remove impurities and dopamine monomers on the surface of the product;

[0035] (5) Disperse the product obtained in step (4) into 100 mmol / L of Ti (SO 4 ) 2 solution, mechanical stirring at room temperature for 2-3 hours;

[0036] (6) The product obtained in step (5) was washed three times with deionized water, and dried in a vacuum d...

Embodiment 2

[0039] Polydopamine modified magnetic spheres and immobilized Ti on the surface 4+ Application of Nanomaterials in Enrichment of Phosphorylated Peptides in β-casein:

[0040] (1) Sample preparation: β-casein in 25mmol / L NH 4 HCO 3 Enzymatic hydrolysis at 37 °C for 16 h in the solution.

[0041](2) Enrichment: 10 μL, 2 mg / mL nanomaterials were dispersed into a 200 μL centrifuge tube containing 0.1% (volume ratio) trifluoroacetic acid and 50% (volume ratio) acetonitrile containing the phosphorylated peptide from step (1), Enriched at 37°C for 30 min; washed with a buffer solution of 0.1% (volume ratio) trifluoroacetic acid and 50% (volume ratio) acetonitrile and magnetically separated 3 times; eluted with 5 μL 0.4 M ammonia water for 10 min.

[0042] (3) Mass spectrometry analysis: Take 0.5 μL of the eluate obtained in step (2) to spot the target and perform mass spectrometry analysis. The mass spectrogram is as follows: Image 6 as shown, Figure 8 It is a polydopamine mo...

Embodiment 3

[0044] Polydopamine modified magnetic spheres and immobilized Ti on the surface 4+ Application of Nanomaterials in Enrichment of Phosphopeptides in Mixed Proteins

[0045] (1) Sample preparation: bovine serum albumin (BSA) was reductively alkylated with dithiothreitol and iodoacetamide, and then enzymatically hydrolyzed at 37 °C for 16 h. β-casein in 25mM NH 4 HCO 3 Enzymatic hydrolysis at 37°C for 16 h in the solution. Add bovine serum albumin (BSA) and β-casein at a molar ratio of 500:1 to a centrifuge tube containing 0.1% (volume ratio) trifluoroacetic acid and 50% (volume ratio) acetonitrile.

[0046] (2) Enrichment: 10 μL, 2 mg / mL The nanomaterial obtained in Example 1 was dispersed into 200 μL containing 0.1% (volume ratio) trifluoroacetic acid and 50% (volume ratio) of the phosphorylated peptide in step (1) In an acetonitrile centrifuge tube, enrich at 37°C for 30 min; wash with a buffer solution of 0.1% (volume ratio) trifluoroacetic acid and 50% (volume ratio) ac...

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Abstract

The invention relates to polydopamine modified magnetic balls, a synthetic method of nanomaterial by fixing Ti<4+> on the surfaces and an application thereof. The synthetic method comprises the following steps: dispersing magnetic balls in a dopamine-contained water solution, quickly adding a trismetyl aminomethane buffer solution in the dopamine-contained water solution in which the magnetic balls are uniformly dispersed under the mechanical stirring condition, mechanically stirring for 10 to 16 hours at a room temperature, and washing and carrying out the magnetic separation to obtain nanomaterial which is good in dispersivity in the water and provided with the polydopamine modified magnetic balls on the surfaces, and dispersing the material in a 100 mmol / L Ti(SO4)2 solution, mechanically stirring for 2 to 3 hours at the room temperature, and washing and carrying out the magnetic separation to obtain polydopamine modified magnetic balls, and nanomaterial by fixing Ti<4+> on the surfaces of the magnetic balls.. The method for fixing Ti<4+> is very simple, a layer of polydopamine is modified on the surface, so that the material has excellent environmental stability and biocompatibility, and is good in dispersivity in the water, and the material can fix a large amount of Ti<4+>, and takes the magnetic balls as a core to cause the material to be simple to operate and carry out magnetic separation, the synthetic method is simple, and low in cost, and can enrich phosphorylated peptide by a high-flux MALDI-TOFMS.

Description

technical field [0001] The invention belongs to the field of advanced nanomaterials and nanotechnology, and specifically relates to a high-throughput MALDI-TOF MS enrichment of phosphorylated peptide polydopamine-modified magnetic balls and fixing Ti on the surface 4+ Synthesis and application of nanomaterials. Background technique [0002] Reversible phosphorylation of protein is a very important post-translational modification, which regulates almost all life activities such as cell signal transduction, cell differentiation, cell growth, cell apoptosis, etc., and is also vividly described as a molecule of cell physiological activities switch. Therefore, the study of protein phosphorylation helps to fully understand the functions and mechanisms of proteins. At present, the research method of protein phosphorylation is to first digest the protein, and then use biological mass spectrometry to accurately identify the amino acid sequence and phosphorylation site of the phosph...

Claims

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Application Information

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IPC IPC(8): H01F1/26C08G73/02G01N1/40G01N27/62
Inventor 邓春晖闫迎华卜静王先颖王梦依孙念荣熊娅
Owner FUDAN UNIV
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