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Nano chitosan derivative affinity matrix and preparation method and application thereof

A technology of chitosan derivatives and nano-chitosan, which is applied in the field of biomedical nanomaterials, can solve impossible problems and achieve stable properties, good biocompatibility, and cheap materials

Inactive Publication Date: 2012-05-30
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The system is easy and time-saving in sample preparation and analysis, and has unique advantages in detecting low-abundance, low-molecular-weight proteins, but identification of adsorbed analytes is impossible

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Preparation of chitosan-GMA epoxy medium (chitosan-GMA, intermediate 1)

[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, 752.4...

Embodiment 2

[0031] Preparation of chitosan-GMA-IDA carboxylic acid medium (chitosan-GMA-IDA, intermediate 2)

[0032] 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.

[0033] 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

[0035] Preparation of Chitosan-GMA-IDA-Cu(II)(chitosan-GMA-IDA-Cu(II)) Affinity Matrix for Immobilizing Transition Metal Ions

[0036] Put the chitosan-GMA-IDA carboxylic acid medium prepared in Example 2 into a beaker, add 20mL concentration of 100mM copper sulfate solution, stir, react at room temperature for 2 hours, filter, wash with water, dry, and grind to obtain a solid powder . The affinity medium is a nanometer material with a particle size of 20-100nm.

[0037] Structure: image 3 C is a transmission electron microscope image of the chitosan-GMA-IDA-Cu(II) affinity medium, as can be seen from the figure, this material is spherical, the size is 20-100nm, and the surface is combined with metal copper ions; Spectral method measures the content of copper to be 25.5mg / g (average value of three times); Its infrared spectrum ( figure 2 .4) The characteristic peaks are: 3423.2, 2927.8, 1729.8, 1633.6, 1510.4, 1452.5, 1386.5, 1253.1, 1154.8, 1122.5, 1065.3, 906.3, 841.1, ...

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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 affinity matrix, a preparation method thereof and a method of applying the affinity matrix to separate and enrich protein molecules. 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 mediumand iminodiacetic acid or aminotriacetic acid or ethylenediamine triacetic acid carry out ring-opening addition reaction to obtain a nano composite medium chitosan derivative with carboxylic active functional groups; and finally the composite medium and a transition metal ion carry out complexing action to prepare the final nano chitosan derivative affinity matrix. The method for separating and enriching protein molecules comprises the steps of adsorbing analytes on the affinity matrix under different adsorption and elution conditions and determining and analyzing the analytes reserved on themedium by mass spectra. The method can be applied to the fields of biology, medicines and environment, including clinical diagnosis and discovery of new medicines.

Description

[0001] A nano-chitosan derivative affinity medium, its preparation method, and a method for separating and enriching protein molecules using the affinity medium technical field [0002] The invention relates to the technical field of biomedical nanometer materials. Specifically, the invention provides a nano-chitosan derivative affinity medium, a preparation method thereof, and a method for separating and enriching protein molecules using the affinity medium. The nano-chitosan derivative affinity medium can specifically adsorb and bind certain protein molecules, so as to separate functional proteins in complex biological systems. Background technique [0003] Proteomics is a new concept proposed by Australian scientist Wilkins in 1994, and it is a new growth point of life science research in the post-genome era. It is to reveal the process of life and explain the mechanism of gene expression regulation through the quantitative study of the overall protein expressed by the g...

Claims

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Application Information

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