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Preparation method and application of macroporous chitosan-polyvinyl alcohol crosslinking affinity membrane chelated with metal ions

A technology of polyvinyl alcohol film and metal ions, which is applied in the interdisciplinary field of biochemistry, can solve the problems of large steric hindrance and hinder the application of chitosan, and achieve the reduction of steric hindrance, high-efficiency affinity adsorption capacity, and energy consumption low effect

Inactive Publication Date: 2015-12-09
WUHAN UNIV OF SCI & TECH +1
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  • Summary
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Proteins are macromolecules, and there is a large steric hindrance when interacting with metal ions on the membrane, which hinders the application of chitosan in affinity chromatography

Method used

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  • Preparation method and application of macroporous chitosan-polyvinyl alcohol crosslinking affinity membrane chelated with metal ions
  • Preparation method and application of macroporous chitosan-polyvinyl alcohol crosslinking affinity membrane chelated with metal ions
  • Preparation method and application of macroporous chitosan-polyvinyl alcohol crosslinking affinity membrane chelated with metal ions

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preparation example Construction

[0017] The embodiment of the present invention provides a kind of preparation method of the macroporous chitosan-polyvinyl alcohol cross-linked affinity film of chelating metal ions, comprising the following steps:

[0018] Step 1: Chitosan with a concentration of 1%-3% and polyvinyl alcohol with a concentration of 3%-9% are mixed in a volume ratio of 1: (2-3), and 200-300 mesh silica gel 2.5%- 7.5%, mix evenly, react in a water bath at 50-60°C for 24-30 hours, pour the reaction mixture solution into a film-making template to obtain a chitosan-polyvinyl alcohol film. Wherein, the chitosan with a concentration of 1%-3% is dissolved in 2-5% acetic acid aqueous solution, and the polyvinyl alcohol with a concentration of 3%-9% is dissolved in water. In this preferred embodiment, chitosan and polyvinyl alcohol are mixed in any ratio of 1:2 to 3:8 by volume.

[0019] Step 2: Soak the chitosan-polyvinyl alcohol membrane with 1-3mol / L sodium hydroxide solution for 1-3 hours to obtain...

specific Embodiment

[0039]

[0040]

[0041] The results of the above experiments show that the prepared macroporous chitosan-polyvinyl alcohol cross-linked affinity membrane connected with iminodiacetic acid (IDA), Cibacronblue3GA, and trimethylolethylenediamine (TED) can Effectively chelate metal ions to avoid the loss of metal ions. It has a large adsorption capacity for histidine-tagged proteins, and has great application prospects in protein separation.

[0042] In summary, the preparation method of the macroporous chitosan-polyvinyl alcohol cross-linked affinity membrane of the chelated metal ion provided by the present invention, the affinity membrane prepared by the above steps has stronger mechanical strength, and is compatible with Compared with the adsorption capacity of the currently prepared membrane to histidine-labeled protein, its advantage is that it can avoid the loss of metal ions, reduce the steric hindrance between the metal ion and the target protein, and increase its ...

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Abstract

The invention relates to a preparation method and application of a macroporous chitosan-polyvinyl alcohol crosslinking affinity membrane chelated with metal ions. The preparation method of the membrane comprises the steps that chitosan, polyvinyl alcohol and silica gel are mixed to prepare a chitosan-polyvinyl alcohol membrane; processing and pore forming are performed with a sodium hydroxide solution; the chitosan-polyvinyl alcohol membrane is added in epoxy chloropropane and sodium hydroxide to obtain a macroporous chitosan-polyvinyl alcohol crosslinking membrane of a net structure; the macroporous chitosan-polyvinyl alcohol crosslinking membrane is steeped in a sodium carbonate solution containing iminodiacetic acid; the dried membrane is steeped in a solution containing divalent metal ions, and then the macroporous chitosan-polyvinyl alcohol crosslinking affinity membrane chelated with the metal ions is obtained. The macroporous chitosan-polyvinyl alcohol crosslinking affinity membrane chelated with the metal ions has the higher mechanical strength and has the efficient affinity adsorption capacity on protein containing histidine; the macroporous chitosan-polyvinyl alcohol crosslinking affinity membrane chelated with the metal ions is applied to separation of protein or enzymes containing the histidine, and the separation method comprises the steps of crude enzyme fluid preparation, specificity affinity adsorption of the membrane on the protein or enzymes containing the histidine, adsorption protein elution and membrane regeneration.

Description

technical field [0001] The invention relates to a preparation method and application of a macroporous chitosan-polyvinyl alcohol cross-linked affinity membrane for chelating metal ions, which belongs to the preparation of new materials and belongs to the field of interdisciplinary biochemistry. Background technique [0002] With the rapid development of related disciplines such as life science and bioengineering, especially the great achievements of genetic engineering, people can produce the desired target products in cells, so the requirements for the separation and purification of biological products are also getting higher and higher. Affinity chromatography is an important method for protein purification, which has high selectivity and separation performance with a large capacity. The affinity chromatography group should be composed of an affinity carrier and an affinity ligand. Traditional carriers and ligands are expensive and not suitable for large-scale production....

Claims

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Application Information

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IPC IPC(8): C08J9/40C08J9/26C08J5/18C08J3/24C08L5/08C08L29/04C08K5/1515C12N9/00C07K1/22
Inventor 曾嵘靳步昆杨忠华阮涛侯亚利龚志伟
Owner WUHAN UNIV OF SCI & TECH
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