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Chitosan and metal copper ion complex western imprinting polymer and its preparation method

A technique for western imprinting and polymer imprinting, which is applied in other chemical processes, chemical instruments and methods, etc., can solve the problems of complex reactions, protein denaturation and inactivation, etc., and achieve simple and controllable operation, low cost, and good reusable effect Effect

Inactive Publication Date: 2011-12-14
WUHAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the prior art of preparing western imprinted polymers, steps such as chemical polymerization are generally required, the reaction is relatively complicated, and it is easy to cause protein denaturation and inactivation during the preparation process.

Method used

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  • Chitosan and metal copper ion complex western imprinting polymer and its preparation method
  • Chitosan and metal copper ion complex western imprinting polymer and its preparation method
  • Chitosan and metal copper ion complex western imprinting polymer and its preparation method

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preparation example Construction

[0035] The concrete steps of above-mentioned preparation method comprise:

[0036] (1) Preparation of chitosan and metal copper ion complexes:

[0037] Weigh 1.5g-3g chitosan, add it into 50mL-100mL 2% acetic acid solution with a mass percentage of 2%, stir until the chitosan is fully dissolved, remove bubbles by ultrasonic to prepare a 3mg / mL chitosan solution, and add it to Add 0.1~0.2gCuSO 4 ·5H 2 O, stirring and reacting for 12 to 24 hours, making the color of the mixture into a clear blue solution;

[0038] (2) Preparation of Western imprinted polymer spherical particles:

[0039] To the solution obtained in (1), add 0.15g ~ 0.3g template protein bovine serum albumin, stir for 12 ~ 24 hours, make it fully dissolved and mix evenly, and then use a medical syringe (7# needle) to inject 2 ~ 4mL Add it dropwise to 1L~2L of 2M sodium hydroxide solution at a rate of 1 / min to obtain white spherical particles. After the white spherical particles settle to the bottom, separate...

Embodiment 1

[0049] Weigh 1.5g chitosan, add it to 50mL mass percentage of 2% acetic acid solution, stir until chitosan is fully dissolved, remove bubbles by ultrasonic to prepare 3mg / mL chitosan solution, and then add 0.1g CuSO 4 ·5H 2 0, stirred and reacted for 12 hours, and the color of the mixture was turned into a clear blue solution; 0.15g template protein bovine serum albumin was added thereto, and after stirring for 12 hours, it was fully dissolved and mixed uniformly, and then used a medical syringe (7# Needle) was added dropwise to 1 L of 2M sodium hydroxide solution at a rate of 3 mL / min to obtain white spherical particles. After the pellets were left standing in the sodium hydroxide solution for 10 minutes, the white spherical particles settled to the bottom, filtered out the white spherical particles and washed with deionized water for 4 times until the pH of the solution reached 7; the western imprinted polymer spherical particles were transferred to In a Erlenmeyer flask wi...

Embodiment 2

[0051] Weigh 3g of chitosan, add it to 100mL of 2% acetic acid solution by mass percentage, stir until the chitosan is fully dissolved, remove bubbles by ultrasonic to prepare a 3mg / mL chitosan solution, and then add 0.2g CuSO 4 ·5H 2 O, stirring and reacting for 24 hours, so that the color of the mixture becomes a clear blue solution; then add 0.3g template protein bovine serum albumin to the obtained solution, stir for 24 hours, fully dissolve and mix evenly, and then use medical Add the syringe (7# needle) dropwise into 2L of 2M sodium hydroxide solution at a rate of 4 mL / min to obtain white spherical particles. After the pellets were left standing in the sodium hydroxide solution for 20 minutes, the white spherical particles precipitated to the bottom, filtered out and washed with deionized water for 6 times until the pH value of the solution reached 7; the resulting western imprinted polymer spherical particles Transfer to the Erlenmeyer flask that 100mL distilled water ...

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Abstract

A chitosan and metal copper ion complex protein imprinting polymer, using the complex formed by natural polymer chitosan and metal copper ion as a functional monomer, imprinting template protein molecules through molecular imprinting technology, and preparing protein selective Western blotting polymers with strong adsorption and separation functions. The polymer also utilizes the interaction between chitosan molecules and template protein molecules, as well as metal copper ions and template protein molecules, effectively increasing the recognition sites for the combination of imprinted polymers and template protein molecules, thereby improving the imprinted polymers. The ability to recognize template protein molecules and the ability to selectively adsorb and separate template protein molecules. The imprinted polymer has good protein selective adsorption and separation effects, so it can be used as a protein-specific selective adsorption and separation material. At the same time, the preparation method of the material is simple and convenient, easy to control, good reusable effect, and low cost.

Description

technical field [0001] The invention relates to the fields of macromolecular chemistry and protein selective adsorption and separation, in particular to a chitosan and metal copper ion complex protein imprinted polymer with protein selective adsorption and separation functions, as well as its preparation method and application. Background technique [0002] In recent decades, protein separation technology has developed rapidly, and people have always hoped to artificially synthesize protein separation media with similar biomolecular affinity recognition capabilities. Molecular imprinting technology is a novel and effective separation technology developed in recent years. At present, the template molecules used in molecular imprinting have developed from traditional small molecules to large molecules such as proteins. The protein molecular imprinting technique for protein separation was developed under this background [Analytica Chimica Acta, 2004, 504: 163-166.]. [0003] I...

Claims

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Application Information

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IPC IPC(8): C08B37/08C08J3/24C08J9/26B01J20/26
Inventor 王艺峰徐敏丁利斌陈红
Owner WUHAN UNIV OF TECH
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