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Preparation method of protein-printed polymer of carboxymethyl chitosan and chitosan

A carboxymethyl chitosan and western blotting technology is applied in the field of preparation of western blotting polymers, which can solve the problems of denaturation inactivation, low separation efficiency, etc., and achieves simple preparation method, mild preparation conditions, and good protein selective separation. effect of function

Inactive Publication Date: 2011-01-12
WUHAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The functional monomers used in traditional molecular imprinting technology are small molecular substances such as acrylic acid and acrylamide. In the presence of template molecules, small molecular functional monomers are polymerized in organic solvents to prepare molecularly imprinted polymers. However, the polymerization process and organic Solvents tend to cause denaturation and inactivation of biological macromolecules such as proteins
In addition, Western blotting polymers still generally have the problem of low selective adsorption and separation efficiency.

Method used

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  • Preparation method of protein-printed polymer of carboxymethyl chitosan and chitosan
  • Preparation method of protein-printed polymer of carboxymethyl chitosan and chitosan
  • Preparation method of protein-printed polymer of carboxymethyl chitosan and chitosan

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The preparation method of the Western imprint polymer of carboxymethyl chitosan and chitosan, it comprises the steps:

[0035] 1) preparation of chitosan derivatives-carboxymethyl chitosan:

[0036] First, weigh 7.5 g of monochloroacetic acid and dissolve it in 20 mL of isopropanol to prepare a carboxymethylation reagent. Then put 4g of NaOH, 16mL of water and 64mL of isopropanol into the reaction vessel, stir well to dissolve, add 5g of chitosan, and stir at 50°C for 1h. Slowly add the carboxymethylation reagent dropwise to the reaction vessel at a rate of 2 mL / min. After continuing the reaction for 4 hours, add 200 mL of 70% ethanol solution by volume to terminate the reaction, then suction filter and wash with 70% ethanol The solution was washed 4 times to obtain a sample; after the obtained sample was vacuum-dried at room temperature for 24 hours, it was placed in 200 mL of ethanol solution with a volume percentage of 80%, and 1 mol / L of HCl was added dropwise ther...

Embodiment 2

[0055] The preparation method of the Western imprint polymer of carboxymethyl chitosan and chitosan, it comprises the steps:

[0056] 1) preparation of chitosan derivatives-carboxymethyl chitosan:

[0057] First, weigh 15 g of monochloroacetic acid and dissolve it in 40 mL of isopropanol to prepare a carboxymethylation reagent. Then put 8g of NaOH, 32mL of water and 128mL of isopropanol into the reaction vessel, stir well to dissolve, add 10g of chitosan, and stir at 60°C for 2h. Slowly add the carboxymethylation reagent dropwise to the reaction vessel at a rate of 4 mL / min. After continuing the reaction for 5 hours, add 300 mL of 90% ethanol solution by volume to terminate the reaction, then filter with suction and wash with 90% ethanol The solution was washed 6 times to obtain a sample. After the obtained sample was vacuum-dried at room temperature for 48 hours, it was placed in 300 mL of ethanol solution with a volume percentage of 90%, and 3 mol / L of HCl was added dropwis...

Embodiment 3

[0068] The preparation method of the Western imprint polymer of carboxymethyl chitosan and chitosan, it comprises the steps:

[0069] 1) preparation of chitosan derivatives-carboxymethyl chitosan:

[0070] First, weigh 7.5 g of monochloroacetic acid and dissolve it in 20 mL of isopropanol to prepare a carboxymethylation reagent. Then put 4g of NaOH, 16mL of water and 64mL of isopropanol into the reaction vessel, stir well to dissolve, add 5g of chitosan, and stir at 55°C for 2h. Slowly add the carboxymethylation reagent dropwise to the reaction vessel at a rate of 4 mL / min. After continuing the reaction for 5 hours, add 300 mL of 80% ethanol solution to terminate the reaction, then filter with suction and wash with 80% ethanol The solution was washed 6 times to obtain a sample. After the obtained sample was vacuum-dried at room temperature for 48 hours, it was placed in 300 mL of ethanol solution with a volume percentage of 90%, and 3 mol / L of HCl was added dropwise therein t...

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Abstract

The invention relates to a preparation method of a protein-printed polymer of carboxymethyl chitosan and chitosan, belonging to the field of polymer chemistry and protein selective adsorption and separation. The preparation method is characterized by comprising the following steps of: (1) preparing carboxymethyl chitosan as a chitosan derivative; (2) preparing a blend solution of the carboxymethyl chitosan and the chitosan; (3) preparing polymer spherical particles embedded with protein; (4) crosslinking the polymer spherical particles embedded with the protein; and (5) eluting template protein to obtain the protein-printed polymer of the carboxymethyl chitosan and the chitosan. The protein-printed polymer of the carboxymethyl chitosan and the chitosan has good capacity of selectively absorbing and separating the template protein and good reusable performance, meanwhile, the preparation method has the advantages of high simplicity, easy control, mild preparation condition and low cost.

Description

technical field [0001] The invention belongs to the field of macromolecular chemistry and protein selective adsorption and separation, in particular to a preparation method of carboxymethyl chitosan and chitosan protein imprinted polymer with functions of protein selective adsorption and separation. Background technique [0002] Protein is an important part of life and the basis of all life activities. To reveal the essence of life activities, it is necessary to study proteins in depth. However, there are tens of thousands of proteins in nature, how to effectively adsorb, separate and purify these proteins has always been a hot and difficult issue that researchers are concerned about. In recent years, with the development of protein separation technology, molecular imprinting technology has been introduced into the field of protein selective adsorption and separation. The functional monomers used in traditional molecular imprinting technology are small molecular substances ...

Claims

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Application Information

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IPC IPC(8): C08J9/26C08J3/24C08J3/12C08L5/08C08B37/08C08K5/1515B01J20/24B01J20/30B01J20/28
Inventor 王艺峰徐敏王立莹刘洲
Owner WUHAN UNIV OF TECH
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