Preparation of gelatine- aminosilane- -anode alumina compound film with affinity

A technique of anodized aluminum and aminosilane, applied in chemical instruments and methods, membrane technology, semipermeable membrane separation, etc. The effect of small mass impedance and improving the efficiency of affinity separation

Inactive Publication Date: 2010-05-19
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The anodized aluminum oxide membrane has ideal pore size and porosity distribution and good chemical and mechanical stability, and has the potential to be used as an affinity membrane matrix membrane. However, due to the low content of active groups on the anodized aluminum oxide membrane, it is difficult to immobilize a large amount of It must be chemically modified to meet the requirements as an affinity separation medium. However, there are no relevant reports on the modification of anodized aluminum oxide membranes.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] 1. Activation of anodic aluminum oxide film: mix 8ml of γ-aminopropyltriethoxysilane, 30ml of absolute ethanol and 3ml of sodium acetate-glacial acetic acid buffer (pH5.0), take a piece with a pore size of 0.15μm and a thickness of An anodic aluminum oxide film with a diameter of 200 μm and a porosity of 85% was placed in the above mixed solution and reacted at 70° C. for 0.5 h. After the film was taken out and washed with ethanol, it was heated at 150°C for 1 hour to obtain an anodic aluminum oxide film with amino groups.

[0017] 2. Preparation of gelatin-aminosilane-anodized aluminum composite film: 160 mg gelatin was dissolved in 20 ml pH3.0 hydrochloric acid solution, and 400 mg EDC·HCl was added to obtain a gelatin hydrochloric acid solution containing EDC·HCl. Take a piece of anodic aluminum oxide film with amino groups prepared in step 1, place it in the above solution, and shake it at 20°C for 36 hours. After the reaction, shake and wash the film with a large a...

Embodiment 2

[0022] 1. Activation of anodized aluminum membrane: Mix 3.5ml γ-aminopropyltrimethoxysilane, 30ml absolute ethanol and 2ml sodium acetate-glacial acetic acid buffer (pH5.0), take a piece with a pore size of 0.7μm and a thickness of An anodic aluminum oxide film with a diameter of 80 μm and a porosity of 55% was placed in the above mixed solution, and reacted at 60° C. for 1 h. After the film was taken out and washed with ethanol, it was heated at 150°C for 1 hour to obtain an anodic aluminum oxide film with amino groups.

[0023] 2. Preparation of gelatin-aminosilane-anodized aluminum composite membrane: Dissolve 40 mg of gelatin in 20 ml of pH 5.0 hydrochloric acid solution, add 100 mg of EDC·HCl to obtain a gelatin hydrochloric acid solution containing EDC·HCl. Take a piece of the anodic aluminum oxide film with amino group prepared in step 1 and place it in the above solution and shake it at 35°C for 30 hours. After the reaction, shake and wash the film with a large amount ...

Embodiment 3

[0026] 1. Activation of the anodic aluminum oxide film: mix 5ml of γ-aminopropylmethyldimethoxysilane, 28ml of absolute ethanol and 2ml of sodium acetate-glacial acetic acid buffer (pH 5.0), take a piece of pore size of 0.4μm, An anodic aluminum oxide film with a thickness of 30 μm and a porosity of 65% was placed in the above mixed solution and reacted at 20° C. for 2 hours. After the film was taken out and washed with ethanol, it was heated at 150°C for 1 hour to obtain an anodic aluminum oxide film with amino groups.

[0027] 2. Preparation of gelatin-aminosilane-anodized aluminum composite film: 200mg gelatin was dissolved in 20ml pH 4.0 hydrochloric acid solution, and 0.25g EDC·HCl was added to obtain a gelatin hydrochloric acid solution containing EDC·HCl. Take a piece of anodic aluminum oxide film with amino groups prepared in step 1, place it in the above solution and shake it at 20°C for 24 hours. After the reaction, shake and wash the film with a large amount of dist...

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Abstract

The invention discloses a method for preparing a compatible gelatin, amino silane and anodic alumina composite membrane, and relates to a method for preparing an affinity membrane used for protein separation. The invention provides the method for preparing the compatible gelatin, amino silane and anodic alumina composite membrane which has more active groups (-NH2 group, -COOH group) as well as better hydrophilicity and biocompatibility, is widely applicable to the affinity separation, and takes the amino silane coupling agent as an activator. An anodic alumina membrane is taken as the basement membrane, which is soaked in amino silane solution for reacting, the membrane is taken out for cleaning and drying, so as to obtain the anodic alumina membrane with epoxy groups, then the obtained anodic alumina membrane with the epoxy groups is soaked in gelatin HCL water solution which contains 1-(3-dimethylamino propyl)-3-ethyl carbodiimide hydrochloride (EDC*HCL) for reacting, and then the composite membrane is taken out for cleaning and drying, thus, the compatible gelatin, amino silane and anodic alumina composite membrane is obtained; or the anodic alumina membrane with amino groups is soaked in carbonyldiimidazole acetone solution for reacting, the membrane is arranged in gelatin water solution for reacting, and then the composite membrane is taken out for cleaning and drying, thus, the compatible gelatin, amino silane and anodic alumina composite membrane is obtained.

Description

technical field [0001] The invention relates to a method for preparing an affinity membrane for protein separation, in particular to a method for preparing an affinity gelatin-aminosilane-anodized aluminum composite membrane. Background technique [0002] The granular adsorbent used in traditional column affinity separation has a "dead-end" pore structure. The binding of the target protein to the ligand is limited by the surface liquid film diffusion, pore diffusion and adsorption kinetics. The main mass transfer resistance is Diffusion from pores. Affinity membrane separation has "through hole" or "convection hole" compared with it, which is only affected by the diffusion and adsorption kinetics of the surface liquid film, eliminating the speed control process of pore diffusion, so that it can be used at a lower operating pressure Rapid separation and purification of target proteins at high and high flow rates, shortening operating time, improving purification efficiency, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D69/12B01D71/02B01D71/70B01D71/74
Inventor 石巍沈雨青葛东涛张其清
Owner XIAMEN UNIV
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