Immobilized metal ion affinity chromatography functionalized hybrid monolithic material, and preparation method and application thereof

A technology of monolithic materials and immobilized metals, which is applied in the field of enrichment of phosphorylated peptides, can solve the problems of low specific surface area, limitation of phosphorylated peptides, cumbersome operation steps, etc., and achieve high specific surface area, high adsorption capacity, and easy operation The effect of simple process

Inactive Publication Date: 2019-12-17
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nanomaterials such as titanium dioxide, mesoporous silicon, magnetic materials, and metal-organic framework (MOF) nanoparticles have shown excellent enrichment specificity and sensitivity in the enrichment of phosphorylated peptides, but these materials need to be prepared by Surface modification introduces specific functional groups, and the operation steps are cumbersome
Polymer monolithic material is also a superior phosphorylated peptide enrichment material, which has the advantages of simple preparation, good permeability and wide pH tolerance range, but its relatively low specific surface area limits its phosphorylation in complex systems. Applications of Peptide Enrichment

Method used

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  • Immobilized metal ion affinity chromatography functionalized hybrid monolithic material, and preparation method and application thereof
  • Immobilized metal ion affinity chromatography functionalized hybrid monolithic material, and preparation method and application thereof
  • Immobilized metal ion affinity chromatography functionalized hybrid monolithic material, and preparation method and application thereof

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Experimental program
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Embodiment 1

[0022] First, add 0.01mmol dodecylamine and 0.015mmol CTAB to 8.5mmol water and 3.5mmol ethanol. After ultrasonic dissolution, put it into an ice bath to lower the temperature of the solution to 0°C. Then, under ice bath, 0.8 mmol TEOS and 0.2 mmol DPTS (the total molar number of Si atoms is 1.0 mmol) were quickly added, and after mixing evenly, the mixed solution was placed in a 40° C. water bath. Then, at 90°C, the phosphoroethyl group was hydrolyzed with 37% concentrated hydrochloric acid for 48 hours, and finally washed with water to neutrality, and vacuum-dried at 60°C for 12 hours to prepare a phosphoric acid-functionalized hybrid monolith.

[0023] Weigh 50mg of carbon material into a centrifuge tube, then add 50mL of 100mg / mL titanium sulfate solution into the centrifuge tube containing the carbon material according to the mass ratio of material to titanium sulfate is 1:100, then place the centrifuge tube on the shaker Shake on the bed for 12 hours. Finally, wash with...

Embodiment 2

[0025]First, add 0.025mmol of dodecylamine and 0.015mmol of CTAB into 8.5mmol of water and 3.5mmol of ethanol. After ultrasonically dissolving, put it into an ice bath to lower the temperature of the solution to 0°C. Then, under ice bath, 0.8 mmol TEOS and 0.2 mmol DPTS (the total molar number of Si atoms is 1.0 mmol) were quickly added, and after mixing evenly, the mixed solution was placed in a 40° C. water bath. Then, at 90°C, the phosphoroethyl group was hydrolyzed with concentrated hydrochloric acid for 48 hours, and finally washed with water to neutrality, and vacuum-dried at 60°C for 12 hours to prepare a phosphoric acid-functionalized hybrid monolith.

[0026] Weigh 50 mg of the hybrid monolith material in a centrifuge tube, then add 50 mL of 100 mg / mL titanium sulfate solution into the centrifuge tube containing the carbon material according to the mass ratio of the material to titanium sulfate as 1:100, and then centrifuge The tube was shaken on a shaker for 12h. Fi...

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Abstract

The invention relates to an immobilized metal ion affinity chromatography (IMAC) functionalized hybrid monolithic material with a high specific surface area, prepared through a sol-gel technology, andan application thereof in phosphorylated peptide enrichment. A hybrid monolithic material containing phosphate functional groups on the surface is prepared through a sol-gel reaction by taking tetraethoxysilane (TEOS) and diethylphosphatoethyltriethoxysilane (DPTS) as precursors, the hybrid monolithic material is hydrolyzed in concentrated hydrochloric acid (HCl) to generate phosphoric acid functional groups on the surface, and the hydrolyzed hybrid monolithic material is chelated with Ti<4+> to obtain the Ti<4+>-IMAC hybrid monolithic material. The Ti<4+>-IMAC hybrid monolithic material canadsorb micromolecular pyridoxal 5-phosphate under the action of metal chelation, and can also be used for carrying out high-sensitivity and high-selectivity enrichment on phosphorylated peptide fragments in beta-casein and human cervical carcinoma cell (HeLa) enzymatic hydrolysate.

Description

technical field [0001] The invention relates to an immobilized metal ion affinity chromatography (IMAC) functionalized hybrid monolithic material with a high specific surface area prepared based on a sol-gel method (sol-gel) and its application in phosphorylated peptide enrichment. The specific preparation process is to first use tetraethoxysilane (TEOS) and diethylphosphorylethyltriethoxysilane (DPTS) as precursors to prepare a surface containing phosphate functional groups through sol-gel reaction. The hybrid monolith is then hydrolyzed in concentrated hydrochloric acid (HCl) to generate phosphoric acid functional groups on the surface, and finally combined with Ti 4+ Chelate to get Ti 4+ -IMAC hybrid monolithic materials. Ti according to the present invention 4+ -IMAC hybrid monoliths have the potential to be applied to the enrichment of phosphorylated peptides in complex biological samples. Background technique [0002] As one of the important tools of signal transdu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/281B01J20/22B01J20/30B01D15/38C07K1/22
CPCB01D15/3804B01J20/22B01J20/281B01J2220/4812B01J2220/52B01J2220/54C07K1/22
Inventor 欧俊杰张海洋马淑娟叶明亮
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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