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Preparation of hydrophilic group modified two-dimensional magnetic nano material and application thereof in glycopeptides large-scale enrichment

A technology of two-dimensional nanomaterials and hydrophilic groups, applied in the field of functionalized magnetic nanomaterials, can solve the problems of poor specificity and selectivity, low enrichment efficiency, etc., and achieves a simple material synthesis method, good enrichment effect and high efficiency. The effect of selective enrichment

Inactive Publication Date: 2020-02-11
BEIJING PROTEOME RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although HILIC-based methods have been adopted in many works, commercially available micron-sized HILIC materials still have problems such as low enrichment efficiency, poor specificity and selectivity.

Method used

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  • Preparation of hydrophilic group modified two-dimensional magnetic nano material and application thereof in glycopeptides large-scale enrichment
  • Preparation of hydrophilic group modified two-dimensional magnetic nano material and application thereof in glycopeptides large-scale enrichment
  • Preparation of hydrophilic group modified two-dimensional magnetic nano material and application thereof in glycopeptides large-scale enrichment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1. Preparation of two-dimensional magnetic nanomaterials modified by hydrophilic groups and enrichment and detection of glycopeptides

[0072] 1. Preparation of two-dimensional magnetic nanomaterials modified by hydrophilic groups

[0073] according to figure 1 The schematic diagram of the synthetic route is shown for the preparation of two-dimensional magnetic nanomaterials modified by hydrophilic groups. The specific steps are as follows:

[0074] Step 1: Preparation of two-dimensional nanomaterial molybdenum disulfide

[0075] 1.21g Na 2 MoO 4 2H 2 O, 1.52g (NH 2 ) 2 CS, and 30mg PEG-20,000 were added to 30mL ultrapure water, mechanically stirred for 30 minutes, and ultrasonically vortexed to obtain a uniform and transparent solution. The resulting solution was transferred to a 50 mL stainless steel autoclave and then sealed, heated and reacted in an electric oven at 220° C. for 20 hours and then cooled to room temperature. The reaction product was ce...

Embodiment 2

[0086] Example 2. Sensitivity detection of two-dimensional magnetic nanomaterials modified with hydrophilic groups for glycopeptide enrichment

[0087] Weigh 1 mg of hydrophilic group-modified two-dimensional magnetic nanomaterials, ultrasonically disperse them in 1 mL of enrichment buffer (anhydrous acetonitrile / water / trifluoroacetic acid, 88:11.9:0.1, v / v / v), and then pipette 40 μL The above suspension was separated from magnetic nanoparticles by applying an external magnetic field, and washed three times with enrichment buffer (anhydrous acetonitrile / water / trifluoroacetic acid, 88:11.9:0.1, v / v / v).

[0088] The above two-dimensional magnetic nanomaterials (40 μg) were resuspended in 50 μL enrichment buffer (anhydrous acetonitrile / water / trifluoroacetic acid, 88:11.9:0.1, v / v / v), and 1 μL (50 pmol / mL or 10pmol / mL) human immunoglobulin enzymatic hydrolysis solution, shake and mix at room temperature for 30min, and then separate the magnetic nanomaterials under the action of a...

Embodiment 3

[0090] Example 3. Selective detection of glycopeptide enrichment by hydrophilic group-modified two-dimensional magnetic nanomaterials

[0091] Weigh 1 mg of hydrophilic group-modified two-dimensional magnetic nanomaterials, ultrasonically disperse them in 1 mL of enrichment buffer (anhydrous acetonitrile / water / trifluoroacetic acid, 88:11.9:0.1, v / v / v), and then pipette 40 μL The above suspension was separated from magnetic nanoparticles by applying an external magnetic field, and washed three times with enrichment buffer (anhydrous acetonitrile / water / trifluoroacetic acid, 88:11.9:0.1, v / v / v).

[0092] The above two-dimensional magnetic nanomaterials (40 μg) were resuspended in 50 μL of enrichment buffer (anhydrous acetonitrile / water / trifluoroacetic acid, 88:11.9:0.1, v / v / v), and 1 μL of human immunoglobulins were added thereto Proteolysis solution and bovine serum albumin enzymatic solution mixture (IgG:BSA 1:200, w / w, 1μg / μL), shaking and mixing at room temperature for 40min,...

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Abstract

The invention discloses a hydrophilic group modified two-dimensional magnetic nano material and a preparation method thereof, and application of a hydrophilic group modified two-dimensional magnetic nano material. The material is prepared from a two-dimensional molybdenum disulfide nano material, magnetic nanoparticles connected to the surface of the two-dimensional molybdenum disulfide nano material through metal covalent bonds, nanogold wires polymerized on the surface of the two-dimensional molybdenum disulfide nano material in an Au-S bond mode, and a hydrophilic group reagent polymerizedon the surface of the nanogold wires in an Au-S bond mode. Through hydrophilic interaction and electrostatic interaction between a group reagent in the material and a glycosylated peptide fragment, efficient selective enrichment of glycopeptides is achieved; and moreover, the glycosylated peptide fragment is good in enrichment effect, and enrichment detection of glycopeptides in low-concentrationhuman serum albumin (such as IgG) and human urine exosome protein can be achieved.

Description

technical field [0001] The invention belongs to the technical field of functionalized magnetic nanomaterials, and in particular relates to the preparation of a hydrophilic group-modified two-dimensional magnetic nanomaterial and its application in large-scale enrichment of glycopeptides. Background technique [0002] Protein glycosylation plays a vital role in many biological processes and is closely related to the occurrence and development of various cancers. Therefore, it is crucial to analyze abnormal changes in glycosylated peptides in complex biological samples. The current research strategy for glycoproteins is mainly to enzymatically digest glycoproteins into peptides, and then analyze them by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) or liquid chromatography tandem mass spectrometry (LC-MS / MS). However, the low abundance of glycopeptides in the tryptic peptide mixture and the overwhelming signal suppression by non-g...

Claims

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Application Information

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IPC IPC(8): G01N1/40G01N30/08B82Y30/00B82Y40/00C01G39/06C01G49/08
CPCB82Y30/00B82Y40/00C01G39/06C01G49/08C01P2004/62C01P2004/64G01N1/405G01N30/08
Inventor 秦伟捷张养军张汉卿
Owner BEIJING PROTEOME RES CENT
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