Nano chitosan derivative and preparation method and application thereof

A technology of nano-chitosan and derivatives, applied in the field of biomedical nano-materials, can solve the problems of enrichment and purification of phosphorylated polypeptides, and achieve good biocompatibility, strong adsorption capacity, and cheap materials

Inactive Publication Date: 2010-06-23
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the enrichment and purification of phosphorylated polypeptides using chitosan derivatives as immobilized substrates

Method used

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  • Nano chitosan derivative and preparation method and application thereof
  • Nano chitosan derivative and preparation method and application thereof
  • Nano chitosan derivative and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: the preparation of chitosan-GMA epoxy medium (chitosan-GMA)

[0028] In a 100mL three-necked flask equipped with a stirrer, a thermometer and a condenser, 0.5g of chitosan (Qingdao Haihui Biological Co., Ltd.) was dissolved in 30mL of an aqueous solution containing dilute acetic acid (2wt%), and 0.5mL of formazan was added. Glycidyl acrylate, stir, then add 0.035 g of ammonium persulfate and 0.035 g of sodium thiosulfate, heat up to 50 ° C, react for 2 hours, stop the reaction, after cooling down to room temperature, centrifuge to remove the supernatant, and then wash with water , to obtain a solid.

[0029] Structure: image 3 A is the chitosan-GMA epoxy dielectric negative staining transmission electron microscope image, as can be seen from the figure, this material is spherical, and the size is 20-100nm; its infrared spectrum ( figure 2 .2) The characteristic peaks are: 3410.2, 2927.2, 1730.7, 1639.8, 1452.3, 1259.8, 1157.9, 1076.9, 905.7, 846.0, 752...

Embodiment 2

[0030] Embodiment 2: the preparation of chitosan-GMA-IDA carboxylic acid medium (chitosan-GMA-IDA)

[0031] In the 100mL three-necked flask equipped with stirrer, thermometer and condenser tube, the composite medium prepared in Example 1 is charged, and 0.5 gram of sodium iminodiacetate, 0.25 gram of sodium chloride and 20 mL of sodium carbonate of 2N are added The solution was heated up to 60° C., reacted for 5 hours, stopped the reaction, cooled to room temperature, filtered, washed with water until neutral, and a solid was obtained.

[0032] Structure: image 3 The negative staining transmission electron microscope figure of B chitosan-GMA-IDA carboxylic acid medium, as can be seen from the figure, this material is spherical, and size is 20-100nm; Its infrared spectrum ( figure 2 .3) The characteristic peaks are: 3437.9, 2929.8, 1929.5, 1640.1, 1607.3, 1452.1, 1387.3, 1253.4, 1154.4, 1072.1, 908.5, 842.1, 754.6; elemental analysis results are C 46.89%, H 7.19%, N 2.46%. ...

Embodiment 3

[0033] Embodiment 3: prepare the chitosan-GMA-IDA-Fe(III) (chitosan-GMA-IDA-Fe(III)) of fixed transition metal ion

[0034] Put the chitosan-GMA-IDA prepared in Example 2 into a beaker, add 20 mL of 100 mM ferric chloride solution, stir, react at room temperature for 2 hours, filter, wash with water, dry, and grind to obtain a solid powder.

[0035] Structure: image 3 C is a transmission electron microscope figure of chitosan-GMA-IDA-Fe(III), as can be seen from the figure, the particle size of this nanomaterial is 20-100nm, and the surface is combined with metal iron ions; The iron content measured by the spectrometer was 15.52 mg / g (average value of three times). Its infrared spectrum ( figure 2 .4) The characteristic peaks are: 3424.0, 2927.2, 1728.8, 1635.0, 1510.4, 1454.5, 1384.7, 1249.7, 1158.1, 1073.1, 908.9, 834.4, 753.0. Elemental analysis: C 43.87%, H 6.93%, N 2.33%.

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Abstract

The invention relates to the technical field of nano materials in biomedicine. Specifically, the invention provides a nano chitosan derivative, a preparation method and a method of applying the nano chitosan derivative to enrich and purify phosphopeptide highly selectively and specifically. The preparation method comprises the following steps: the chitosan is dissolved in diluted acid and then is mixed with glycidyl methacrylate or glycidyl acrylate to carry out self-polymerization and grafting reaction, thus obtaining a nano composite medium chitosan derivative with active epoxy groups; the composite medium and iminodiacetic acid or aminotriacetic acid or ethylenediamine triacetic acid carry out ring-opening addition reaction to obtain a nano medium chitosan derivative with carboxylic active functional groups; and finally the medium and transition metal ions such as Fe3+, Ti4+, Zr4+, Ga3+ and the like carry out chelation to prepare the final nano chitosan derivative. The nano chitosan derivative has high specificity and can be used for enriching and purifying phosphopeptide with low abundance in the biological samples, thus being used in the fields of biology and medicines, including clinical diagnosis.

Description

technical field [0001] The invention relates to the technical field of biomedical nanometer materials. Specifically, the present invention provides a nano-chitosan derivative, a preparation method thereof and a method for using the nano-chitosan derivative to enrich and purify phosphorylated polypeptide with high selectivity and specificity. Background technique [0002] Post-translational protein modification is a hot research topic in proteomics. Protein phosphorylation is the most common and important post-translational modification of proteins. About 30% of proteins encoded by human genes can be phosphorylated. Protein phosphorylation and dephosphorylation almost regulate life In the whole process of activity, including cell proliferation, development and differentiation, neural activity, muscle contraction, metabolism, tumorigenesis, etc., protein phosphorylation is still the main signal transmission mode. [0003] Traditional methods for protein phosphorylation analy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08C08F251/00C08F8/00C08F8/42B01J20/24B01J20/30C07K1/22
Inventor 邹霞娟刘丹钟丽君娄雅欣杨彬彭嘉柔
Owner PEKING UNIV
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