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Nylon affinity membrane preparing method and use

An affinity membrane and nylon membrane technology, applied in the application field of separation and purification of papain, can solve the problems of inability to reach the pharmaceutical industry, long separation time, low separation speed, etc., and achieve large-scale purification separation and easy large-scale purification. The effect of low probability of separation and denaturation

Inactive Publication Date: 2007-03-28
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the purification of papain is by precipitation method, but this method is still mixed with other proteases [6], which cannot meet the needs of the pharmaceutical industry
[0003] As people put forward further requirements for purity and safety, chromatographic techniques are applied to the purification of papain, such as ion exchange chromatography and affinity chromatography [7-9]. Limited by intergranular diffusion and low axial velocity, the pressure drop is large, the separation time is long, the processing volume is small, and high feeding speed cannot be used
Membrane separation is a separation technology based on the size of molecules. It is characterized by large surface area, short diffusion path, low operating pressure, and large processing capacity. However, traditional membrane separation technology can only separate species with a relative molecular mass difference of more than 10 times. Meet the needs of biochemical separation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The nylon membrane was first rehydrolyzed in 0.1M hydrochloric acid for 72h, and then activated with 10% glutaraldehyde. The activated nylon membrane was then placed in a 0.5% chitosan solution for chemical bonding, and then dried in an oven at 70°C , the chitosan-modified nylon membrane was obtained. The content of chitosan bound on the nylon membrane was tested according to the method disclosed in the document [10], and the test result showed that the content of chitosan was 89.6 mg / g membrane.

[0025] Put 2g of chitosan-modified nylon membrane in 50ml of CibacronBlue F3GA dye solution with a concentration of 5.0mg / ml, react at 60°C for 0.5h, then add 20ml of 20% NaCl solution, continue to react for 1.0h, and then add 5.0ml of NaCO3 solution with a concentration of 20% was used for color-fixing reaction. After 2 hours of reaction, it was taken out, washed with hot water, methanol, 2M NaCl, 6M urea, and deionized water in sequence, and stored at 4°C for later use.

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Embodiment 2

[0029] The nylon membrane was re-hydrolyzed in 10M hydrochloric acid for 10h, and then activated with 25% glutaraldehyde. The activated nylon membrane was then placed in a 5% chitosan solution for chemical bonding, and then dried in an oven at 120°C. That is, a chitosan-modified nylon membrane is obtained. The content of chitosan bound on the nylon membrane was tested according to the method disclosed in the literature [10], and the test result showed that the content of chitosan was 90.6 mg / g membrane.

[0030] Put 10g of chitosan modified nylon membrane in 350ml of CibacronBlue F3GA dye solution with a concentration of 30mg / ml, react at 60°C for 3h, then add 50ml of 20% NaCl solution, continue to react for 3.0h, and then add 20ml of concentration The color-fixing reaction was carried out for 20% NaCO3 solution. After 10 hours of reaction, it was taken out, washed with hot water, methanol, 2M NaCl, 6M urea, and deionized water in sequence, and stored at 4°C for later use.

...

Embodiment 3

[0034] The nylon membrane was first rehydrolyzed in 5M hydrochloric acid for 50h, and then activated with 15% glutaraldehyde. The activated nylon membrane was then placed in a 3% chitosan solution for chemical bonding, and then dried in an oven at 100 °C. A chitosan-modified nylon membrane is obtained. The content of chitosan bound on the nylon membrane was tested according to the method disclosed in the literature [10], and the test result showed that the content of chitosan was 89.2 mg / g membrane.

[0035] Put 5g of chitosan-modified nylon membrane in 200ml of CibacronBlue F3GA dye solution with a concentration of 20mg / ml, react at 60°C for 2h, then add 30ml of 20% NaCl solution, continue to react for 2h, and then add 10ml of 20% NaCO3 solution was used for color-fixing reaction. After 5 hours of reaction, it was taken out, washed with hot water, methanol, 2M NaCl, 6M urea, and deionized water in sequence, and stored at 4°C for later use.

[0036] The separation and purific...

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Abstract

The present invention discloses a preparation method of nylon affinity membrane. Said method includes the following steps:(1) on the nylon membrane chemically-bonding chitosan so as to obtain the chitosan modified nylon membrane; and (2) bonding the chitosan modified nylon membrane and affinity dye Cibacron Blue F3GA so as to obtain the described nylon affinity membrane which can be used for separating and purifying papain.

Description

technical field [0001] The invention relates to a preparation method and application of a nylon affinity membrane, in particular to a preparation method of an affinity membrane with nylon as a matrix and dye Cibacron Blue F3GA as an affinity group and its application in separating and purifying papain. application. Background technique [0002] Papain (EC3.4.22.2) is a class of sulfhydryl protease, widely present in the roots, stems, leaves and fruits of papaya (Caricapapaya), among which the most abundant in immature milk [1-2]. It is widely used in the food industry, such as beer clarification and meat tenderization, as well as leather, textile, daily chemical and pharmaceutical industries [3-5]. Most of the purification of papain is by the precipitation method, but this method is still mixed with other proteases [6], which cannot meet the needs of the pharmaceutical industry. [0003] As people put forward further requirements for purity and safety, chromatographic tech...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D71/56C12N9/50
Inventor 朱利民聂华丽
Owner DONGHUA UNIV
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