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Nylon affinity membrane preparing method and use

A technology of affinity membrane and nylon membrane, which is applied in the application field of separation and purification of papain, can solve the problems of not being able to reach the pharmaceutical industry, long separation time, and mixing, and achieve large-scale purification and separation, and easy large-scale purification and separation , the effect of short analysis time

Inactive Publication Date: 2008-12-31
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 hydrolyzed in 0.1M hydrochloric acid for 72h, and then activated with 10% glutaraldehyde. The nylon membrane activated by glutaraldehyde was then placed in 0.5% chitosan solution for chemical bonding, and then heated at 70°C. drying in an oven to obtain a chitosan-modified nylon membrane. The content of chitosan bonded on the nylon membrane was tested according to the method disclosed in literature [10]. The test result showed that the content of chitosan was 89.6mg / g membrane.

[0025] The chitosan-modified nylon membrane 2g that step (1) obtained is placed in the 50mlCibacron Blue F3GA dye solution that concentration is 5.0mg / ml, 60 ℃ reacts 0.5h, then adding 20ml concentration is 20%NaCl solution, continues reaction 1.0h, then add 5.0ml of 20% NaCO 3 The solution was subjected to color fixation reaction, and after 2 hours of reaction, it was taken out, washed with hot water, methanol, 2M NaCl, 6M urea, and deionized water in sequence, an...

Embodiment 2

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

[0030] The chitosan-modified nylon membrane 10g that step (1) obtained is placed in the 350ml Cibacron Blue F3GA dye solution that concentration is 30mg / ml, 60 ℃ of reaction 3h, then add 50ml concentration and be 20%NaCl solution, continue reaction 3.0h , and then add 20ml of 20% NaCO 3 The solution was subjected to a color fixation reaction, and after 10 hours of reaction, it was taken out, washed with hot water, methanol, 2M NaCl, 6M urea,...

Embodiment 3

[0034] The nylon membrane was first hydrolyzed in 5M hydrochloric acid for 50 hours, and then activated with 15% glutaraldehyde. The nylon membrane activated by glutaraldehyde was then placed in a 3% chitosan solution for chemical bonding, and then heated in an oven at 100°C. drying in medium temperature to obtain the chitosan-modified nylon membrane. The content of chitosan bonded on the nylon membrane was tested according to the method disclosed in literature [10]. The test result showed that the content of chitosan was 89.2mg / g membrane.

[0035] The chitosan-modified nylon membrane 5g that step (1) obtained is placed in the 200mlCibacron Blue F3GA dye solution that concentration is 20mg / ml, and 60 ℃ of reaction 2h, add 30ml concentration then and be 20%NaCl solution, continue reaction 2h, Then add 10ml of 20% NaCO 3 The solution was subjected to color fixation reaction, and after 5 hours of reaction, it was taken out, washed with hot water, methanol, 2M NaCl, 6M urea, and...

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Abstract

The invention discloses a method for preparing a nylon affinity membrane, which comprises the following steps: (1) chemically bonding chitosan on the nylon membrane to reduce the non-specific adsorption of the nylon membrane to protein, and obtain chitosan-modified Nylon membrane; (2) Bonding the nylon membrane after chitosan modification treatment and affinity group dye Cibacron Blue F3GA to obtain the nylon affinity membrane; the nylon affinity membrane is used for separating and purifying papain. The beneficial effects of the present invention are: compared with traditional membrane separation and affinity chromatography, the nylon affinity membrane prepared by the present invention not only has high purification ratio, small pressure drop, short analysis time, and the possibility of denaturation of biological macromolecules during the separation process It is small, allows faster feeding speed, and is easier to achieve large-scale purification and separation than column affinity chromatography.

Description

technical field [0001] The present 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 substrate and dye Cibacron Blue F3GA as an affinity base and its application in the separation and purification of papain. application. Background technique [0002] Papain (EC3.4.22.2) is a kind of sulfhydryl protease, which widely exists in the roots, stems, leaves and fruits of papaya (Caricapapaya), and is 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]. The purification of papain mostly adopts 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, chroma...

Claims

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Application Information

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