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Titanium ion fixation affinity chromatography material and preparation and use thereof

A chromatographic material, titanium ion technology, applied in the preparation method of peptide, solid adsorbent liquid separation, separation method, etc., to achieve the effect of high specificity, selectivity and high specificity

Active Publication Date: 2009-04-01
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation and application of immobilized titanium ion affinity chromatography materials have not been reported in the literature before.

Method used

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  • Titanium ion fixation affinity chromatography material and preparation and use thereof
  • Titanium ion fixation affinity chromatography material and preparation and use thereof
  • Titanium ion fixation affinity chromatography material and preparation and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1. Preparation of a polymer affinity material for immobilizing titanium ions

[0041] A polymer with a phosphoric acid group is generated by polymerizing a monomer with a phosphoric acid group and a cross-linking agent, and the polymer is then reacted with a titanium ion solution to obtain a polymer affinity material for immobilizing titanium ions, which is further applied to enrichment and purified phosphopeptides. In order to prepare polymer materials with phosphoric acid groups, it is first necessary to prepare a polymer reaction solution. Monomer and cross-linking agent are the main components of the reaction solution, and they form a polymer skeleton after polymerization. In order to adjust the physicochemical properties of the polymer, various monomers can be used, but one of them must include a monomer with a phosphoric acid group. Porous materials have a greater surface area and therefore a higher adsorption capacity. In order to prepare porous polyme...

Embodiment 2

[0047] Embodiment 2. Preparation of inorganic affinity material for immobilizing titanium ions

[0048] MCM-41 is an inorganic medium that uses a hydrothermal molecular self-organization method, that is, a hexagonal liquid crystal weave structure formed by the interaction of a certain concentration of organic guides (surfactants) and inorganic species (monomers or oligomers). porous silicon material. When the organic directing agent is removed by heat treatment or chemical means, the resulting solid is called MCM-41 mesoporous molecular sieve. The MCM-41 mesoporous molecular sieve presents a multi-level ordered structure, and can have a specific ordered structure or morphology at multiple scales (such as nanoscale, microscale or macroscale). On the nanometer level, MCM-41 has an ordered "honeycomb" porous structure, that is, a one-dimensional linear channel or a hexagonal close-packed array. The pore size can be adjusted within the range of 1.5-30nm, and the most typical pore...

Embodiment 3

[0057] Example 3. Immobilized titanium ion affinity chromatography material for selective enrichment of phosphopeptides

[0058] Preparation of sample solution: 1 mg of α-casein and β-casein were dissolved in 1 mL of 50 mM ammonium bicarbonate solution (pH 8.2), and trypsin was added according to the mass ratio of 40:1 to trypsin The enzymatic hydrolysis reaction was carried out, the reaction time was 16 hours, and the enzymatic hydrolysis temperature was controlled at 37°C. The obtained proteolysis solution was stored in a -30°C refrigerator for future use. 6.6mg of bovine serum albumin and 4.5mg of ovalbumin were dissolved in 1mL of reducing solution (pH8.2, 8M urea, 50mM ammonium bicarbonate solution), placed at room temperature for 4h, and then added to each tube of solution Add 25 μL, 1M DTT solution, reduce at 37°C for 2 hours, then add 50 μL, 1M IAA solution to each tube of solution, let stand in the dark at room temperature for 30 minutes, then dilute ten times with 5...

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Abstract

The invention provides a fixed titanium ion affinity chromatographic material and the preparation and the application thereof. The strong mutual action between a titanium ion and a phosphate group on a phosphoric acid modified solid phase carrier is used for fixing the titanium ion on the carrier. The fixed titanium ion affinity chromatographic material which is enriched in phosphopeptide is used by the invention. Due to the strong chelating function between the fixed titanium ion and the phosphopeptide, the phosphopeptide is kept on the affinity chromatographic material to be separated from the fixed titanium ion.

Description

technical field [0001] The present invention relates to the separation and enrichment of phosphopeptides, specifically a fixed titanium ion affinity chromatography material and its preparation and application in the enrichment of phosphopeptides. The interaction immobilizes the titanium ions on the phosphoric acid-modified solid-phase support to form an immobilized titanium ion affinity chromatography material. Phosphopeptides are retained on the immobilized titanium ion affinity chromatography material due to the strong interaction between the phosphate group and the immobilized titanium ions, thereby achieving specific separation and enrichment of phosphopeptides from complex proteolysis solutions. Background technique [0002] With the completion of the Human Genome Project, life science has entered the era of functional genome. About 2% of the genes in the human genome encode more than 500 protein kinases and more than 100 phosphatases, corresponding to the phosphorylat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/281B01J20/286
CPCB01D15/3828B01J20/3265B01J20/286C07K1/22
Inventor 邹汉法周厚江叶明亮
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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