Synthesis method and application of polydopamine modified graphene nanometer material with Ti<4+> fixed on surface

A nanomaterial, polydopamine technology, applied in the analysis of materials, material analysis by electromagnetic means, preparation of test samples, etc., can solve problems such as reducing the specificity of enriched phosphorylated peptides, and achieve excellent environmental stability and biological stability. Compatibility, High Sensitivity, High Sensitivity Effects

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

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Problems solved by technology

However, the loss of metal ions and severe non-specific adsorption of acidic peptides due to relatively weak interactions during injection and washing greatly reduced their specificity for enriching phosphopeptides.

Method used

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  • Synthesis method and application of polydopamine modified graphene nanometer material with Ti&lt;4+&gt; fixed on surface
  • Synthesis method and application of polydopamine modified graphene nanometer material with Ti&lt;4+&gt; fixed on surface
  • Synthesis method and application of polydopamine modified graphene nanometer material with Ti&lt;4+&gt; fixed on surface

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Experimental program
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Effect test

Embodiment 1

[0032] Polydopamine modified graphene and immobilized Ti on the surface 4+ Synthesis of Nanomaterials:

[0033] (1) Disperse graphene in concentrated nitric acid, stir at 60 °C for 7 hours, wash with water until neutral, and dry in a vacuum oven;

[0034] (2) Disperse 10 mg of acidified graphene into 40 mL of dopamine aqueous solution with a concentration of 1 mg / mL, and make it uniformly dispersed;

[0035] (3) Under the condition of continuous magnetic stirring, quickly add 10 mL concentration of tris buffered solution of 10 mmol / L (pH is 8.5) to the product obtained in step (2);

[0036] (4) The solution obtained in step (3) was stirred and reacted by magnetic force at room temperature for 16 hours;

[0037] (5) washing the product obtained in step (4) three times with deionized water to remove impurities and dopamine monomers on the surface of the product;

[0038] (6) Disperse the product obtained in step (5) into 100 mmol / L of Ti (SO 4 ) 2 solution, magnetically st...

Embodiment 2

[0042] Polydopamine modified graphene and immobilized Ti on the surface 4+ Application of Nanomaterials in Enrichment of Phosphorylated Peptides in β-casein:

[0043] (1) Sample preparation: β-casein in 25mmol / L NH 4 HCO 3 in solution 37 o C enzymatic hydrolysis for 16 h.

[0044] (2) Enrichment: Disperse 10 μL, 2 mg / mL nanomaterials into 200 μL 0.1% volume ratio TFA50% volume ratio ACN centrifuge tube containing the phosphorylated peptide from step (1), enrich at 37°C for 30 min; 0.1% volume ratio TFA and 50% volume ratio ACN buffer wash and magnetic separation 3 times; 5 μL 0.4 M ammonia water elution for 10 min.

[0045] (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: Figure 4 as shown, Image 6 is polydopamine modified graphene and immobilized Ti on the surface 4+ Nanomaterial enrichment of phosphorylated peptides in β-casein with detectio...

Embodiment 3

[0047] Polydopamine modified graphene and immobilized Ti on the surface 4+ Application of Nanomaterials in Enrichment of Phosphopeptides in Mixed Proteins

[0048] (1) Sample preparation: bovine serum albumin (BSA) was first reductively alkylated with dithiothreitol and iodoacetamide, and then at 37 o C enzymatic hydrolysis for 16 h. β-casein in 25mM NH 4 HCO 3 in solution 37 o C enzymatic hydrolysis for 16 h. Add bovine serum albumin (BSA) and β-casein at a molar ratio of 1000:1 to a centrifuge tube containing 0.1% TFA and 50% ACN.

[0049] (2) Enrichment: 10 μL, 2 mg / mL The nanomaterials obtained in Example 1 were dispersed into 200 μL 0.1% volume ratio TFA 50% volume ratio ACN centrifuge tube containing the phosphorylated peptide in step (1), at 37°C Enriched for 30 min; washed with 0.1% volume ratio TFA and 50% volume ratio ACN buffer and magnetically separated 3 times; 5 μL 0.4 M ammonia water was eluted for 10 min.

[0050] (3) Mass spectrometry analysis: Take 0....

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Abstract

The invention relates to a synthesis method and application of a polydopamine modified graphene nanometer material with Ti<4+> fixed on surface. The method comprises the following steps of: at first, dispersing graphene into dopamine-containing aqueous solution, quickly adding trismetyl aminomethane buffer solution into the dopamine-containing aqueous solution with uniformly distributed graphene under magnetic stirring, magnetically stirring at room temperature for 10 hours, washing and centrifugally separating so as to obtain a graphene nanometer material which has good water dispersibility and is modified by dopamine; and then, dispersing the material into 100mmol / L Ti(SO4)2 solution, magnetically stirring at room temperature for 2 hours, washing and centrifugally separating so as to obtain the polydopamine modified graphene nanometer material with Ti<4+> fixed on the surface. The material has the advantages that a method for fixing Ti<4+> is very simple; the surface is modified by a layer of polydopamine, so that the material has excellent environment stability and biocompatibility as well as good water dispersibility; the material can fix a great mount of Ti<4+>; and the synthesis method is simple and low in cost, and phosphrylation peptides are enriched in the matrix-assisted laser desorption / ionization time of a flight mass spectrometry (MALDI-TOFMS).

Description

technical field [0001] The invention belongs to the field of advanced nanomaterials and nanotechnology, in particular to a kind of MALDI-TOF MS enriching phosphorylated peptide polydopamine modified graphene and immobilizing 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 phosphorylated peptide. H...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G83/00G01N27/62G01N1/40
Inventor 邓春晖闫迎华
Owner FUDAN UNIV
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