Magnetic anion exchanger and application of magnetic anion exchanger as endotoxin adsorption medium

An anion exchanger and endotoxin technology, applied in the field of biomedical materials, can solve the problems of limited processing capacity, cumbersome operation, decreased yield, etc., and achieve the effect of high removal rate and high retention rate

Active Publication Date: 2012-04-25
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if the target protein or other biological products in the sample are also negatively charged, the currently commonly used anion exchangers will often adsorb it and cause problems such as a decrease in yield; and ion exchange is often adsorbed by column packing, and the processing capacity is limited to a certain extent , and the operation is relatively cumbersome

Method used

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  • Magnetic anion exchanger and application of magnetic anion exchanger as endotoxin adsorption medium
  • Magnetic anion exchanger and application of magnetic anion exchanger as endotoxin adsorption medium
  • Magnetic anion exchanger and application of magnetic anion exchanger as endotoxin adsorption medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: the preparation of magnetic anion exchanger

[0021] (1) Ultrasonic dispersion of 1.5g ferroferric oxide in 100mL of 2% acetic acid solution by volume fraction, adding 3.0g chitosan (produced by Aoxing Company, molecular weight 300,000, deacetylation degree 95%, i.e. amino density is 5833μmol / g) to make it completely dissolved and then dispersed into 100mL of liquid paraffin containing 1.0mL Span 80, mechanically stirred for 30min, then added dropwise 6.0mL of 40% formaldehyde solution, reacted at 40°C for 2h, added 50mL of petroleum ether, and stood to separate layer, pour out the upper oil phase, add 100mL of distilled water containing 1.0mL Tween 80, stir and adjust the pH to 13 with 10% NaOH solution, while raising the temperature to 70°C, add 4.0mL of epichlorohydrin after reacting for 2h , continue to stir the reaction for 3h, and the product is washed with ethanol and distilled water and dried in vacuum.

[0022] (2) Soak 1.0 g of microspheres obta...

Embodiment 2

[0026] Example 2: Application of Magnetic Anion Exchanger in Adsorbing Endotoxin in Acidic Protein

[0027] Take 25 mg of magnetic anion exchanger, wash with 1% sodium deoxycholate solution and filter three times, then wash three times with pyrogen-free water, dry and disperse into 2.0 ml solution containing 2.0 mg bovine serum albumin (BSA) and 18 EU endotoxin , shake and adsorb for 60 min, absorb the adsorption medium with a magnet, take out the supernatant to determine the protein and endotoxin content.

[0028] The anion exchanger DEAE-Sepharose was used as the adsorption medium, and the adsorption conditions were the same as above, as a comparative experiment.

Embodiment 3

[0029] Example 3: Application of Magnetic Anion Exchanger in Adsorbing Endotoxin in Basic Protein

[0030] Take 25 mg of magnetic anion exchanger, wash with 1% sodium deoxycholate solution and filter three times, then wash three times with pyrogen-free water, dry and disperse into 2.0 ml of a solution containing 1.0 mg of lysozyme (LYZ) and 23 EU of endotoxin, shake Adsorb for 60 minutes, absorb the adsorption medium with a magnet, take out the supernatant to measure the protein and endotoxin content.

[0031] The anion exchanger DEAE-Sepharose was used as the adsorption medium, and the adsorption conditions were the same as above, as a comparative experiment.

[0032] Coomassie Brilliant Blue method was used for the detection of protein content, and the chromogenic matrix method of Limulus reagent was used for the determination of endotoxin content. Figure 4shown. As can be seen from the figure, whether it is an acidic protein BSA solution or a basic protein LYZ solution, ...

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Abstract

The invention discloses a magnetic anion exchanger and an application of the magnetic anion exchanger as an endotoxin adsorption medium. The magnetic anion exchanger is prepared by the following steps: (1) dispersing a magnetic material in an acetic acid solution containing chitosan, then transferring the obtained water phase I to liquid paraffin containing Span 80, and preparing magnetic chitosan microballoons through an inverse suspension method by using formaldehyde and epoxy chloropropane as cross-linking agents; and (2) successively immersing the magnetic chitosan microballoons obtained in the step (1) in an HCl solution and an NaOH solution, after filtering, washing with ethanol, and drying to obtain cross-linked magnetic chitosan microballoons, that is, the magnetic anion exchanger. The magnetic anion exchanger has high endotoxin removal rate, and high protein reservation ratio. According to the invention, because of the introduction of the magnetic material, the operation is convenient, and the magnetic anion exchanger is suitable for removing endotoxin in biological products.

Description

1. Technical field [0001] The invention relates to the field of biomedical materials, in particular to a magnetic anion exchanger and its application as an endotoxin adsorption medium. 2. Background technology [0002] Endotoxin is the general term for toxic substances present in the cells of Gram-negative bacteria. It is a unique structure on the outer membrane of the cell wall of various Gram-negative bacteria. It is called "pyrogen", its main component is lipopolysaccharide (LPS), and its chemical structure is mainly divided into two parts: polysaccharide and lipid A (Lipid A), of which lipid A is the active center of endotoxin. LPS widely exists in drinking water, pharmaceutical preparations, food, and body fluids, and has strong pyrogenicity. A small amount of LPS (2ng / kg) intravenous injection can cause fever, and a large dose can cause blood circulation disorders and LPS shock. Therefore, the removal of LPS has been a research hotspot. At present, the commonly used ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J41/12B01J20/24B01J41/13
Inventor 应国清来灿林易喻梅建凤王鸿陈建澍姜玉奇
Owner ZHEJIANG UNIV OF TECH
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