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Method for preparing layer-by-layer self-assembled protein-imprinted polymer of chitosan

A technology of western imprinting and layer-by-layer self-assembly, which is applied in the fields of polymer chemistry and protein selective adsorption and separation, can solve the problems of complex preparation process, protein denaturation and inactivation, and achieve simple preparation method, reduced denaturation, and good protein selection The effect of adsorption and separation functions

Inactive Publication Date: 2010-07-28
WUHAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] As mentioned above, in the prior art of preparing western imprinted polymers, there are problems such as complicated preparation process and easy denaturation and inactivation of proteins

Method used

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  • Method for preparing layer-by-layer self-assembled protein-imprinted polymer of chitosan
  • Method for preparing layer-by-layer self-assembled protein-imprinted polymer of chitosan
  • Method for preparing layer-by-layer self-assembled protein-imprinted polymer of chitosan

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A preparation method of layer-by-layer self-assembled western imprinted polymer of chitosan, it comprises the steps:

[0030] 1) Preparation of cross-linked chitosan spherical particle substrate:

[0031]① Weigh 1.5g of chitosan, add it to 50mL of 2% acetic acid solution by mass (2% by mass means 2g of acetic acid in 100g of acetic acid solution), stir until the chitosan is fully dissolved, and remove bubbles by ultrasonic Obtain the chitosan solution of 3mg / mL; Add dropwise in the 2mol / L sodium hydroxide solution of 2L with medical syringe (7# needle head) with the drop rate of 2mL / min, white spherical particle appears in the solution, wait until white After the spherical particles settled to the bottom, the white spherical particles were separated and washed repeatedly with deionized water for 6 times until the pH value of the solution reached 7; then separated to obtain spherical chitosan particles;

[0032] ② The separated chitosan particles were transferred to a c...

Embodiment 2

[0043] A preparation method of layer-by-layer self-assembled western imprinted polymer of chitosan, it comprises the steps:

[0044] 1) Preparation of cross-linked chitosan spherical particle substrate:

[0045] 1. Weigh 3.0g chitosan, add it to 100mL of 2% acetic acid solution by mass, stir until chitosan is fully dissolved, and ultrasonically remove air bubbles to obtain a 3mg / mL chitosan solution; then use a medical syringe ( 7# needle) was added dropwise to 2L of 2mol / L sodium hydroxide solution at a rate of 4mL / min, and white spherical particles appeared in the solution. After the white spherical particles settled to the bottom, the white spherical particles were separated and deionized Water was repeatedly washed 6 times until the pH value of the solution reached 7; then separated to obtain chitosan spherical particles;

[0046] ② The separated chitosan spherical particles were transferred to a conical flask filled with 150mL deionized water, and the pH value of the sol...

Embodiment 3

[0057] A preparation method of layer-by-layer self-assembled western imprinted polymer of chitosan, it comprises the steps:

[0058] 1) Preparation of cross-linked chitosan spherical particle substrate:

[0059] 1. Take by weighing 1.5g chitosan, join in the mass percentage of 50mL and be 2% acetic acid solution, after stirring until chitosan is fully dissolved, obtain the chitosan solution of 3mg / mL after ultrasonically removing air bubbles; Then use medical syringe ( 7# needle) was added dropwise to 1L of 2mol / L sodium hydroxide solution at a rate of 2mL / min, and white spherical particles appeared in the solution. After the white spherical particles settled to the bottom, the white spherical particles were separated and deionized Water was repeatedly washed 4 times until the pH value of the solution reached 7; then separated to obtain spherical chitosan particles;

[0060] ② The separated chitosan spherical particles were transferred to a conical flask filled with 50mL deio...

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Abstract

The invention relates to a method for preparing layer-by-layer self-assembled protein-imprinted polymer of chitosan. A cross-linked chitosan spherical particle substrate is first prepared, electronegative template protein BSA and electropositive chitosan are alternately adsorbed by the method of layer-by-layer self-assembly, that is, the template protein and the chitosan are alternately electrostatically self-assembled on the surface of the cross-linked chitosan spherical particle substrate, the imprinting of template protein molecules is carried out, eluent is then adopted to remove the template protein molecules, so that a site and hole structure which can identify the template protein and vice versa is left on the surface of the substrate, consequently, the substrate has the selective adsorption and separation functions of the protein, and the layer-by-layer self-assembled protein-imprinted polymer of chitosan is obtained by the method. The polymer prepared by the method has good capability to selectively adsorb the template protein, and has a good selective protein separation function, meanwhile, the preparation method is simple, preparation conditions are mild, and the method has good recyclability.

Description

technical field [0001] The invention belongs to the fields of macromolecular chemistry and protein selective adsorption and separation, in particular to a preparation method of chitosan layer-by-layer self-assembled protein imprinted polymer with protein selective adsorption and separation functions. Background technique [0002] At present, the efficient and selective adsorption separation of biological macromolecules, especially protein molecules, is a challenging research field that has attracted great attention from researchers. In recent years, the protein molecular imprinting method has become a novel and effective protein selective adsorption separation technology, showing important application value and broad development prospects. At present, molecular imprinting techniques are used to separate proteins mainly include: embedding method, surface imprinting method and epitope method [Chemical Bulletin, 2005, 7: 504-509]. However, there are still some problems in thes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J9/00C08J9/26C08J3/24C08J3/14C08L5/08C08K5/1515
Inventor 王艺峰徐敏陈艳军王立莹
Owner WUHAN UNIV OF TECH
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