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Multifunctional separation medium with tetrazole as functional group and preparing method thereof

A separation medium and multi-purpose technology, applied in the field of chromatographic separation, can solve the problems of metal ion loss and single use, and achieve the effect of fast separation speed, high purity and easy regeneration

Inactive Publication Date: 2008-07-23
NORTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, these ligands still have the risk of metal ion loss, and they have a single purpose

Method used

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  • Multifunctional separation medium with tetrazole as functional group and preparing method thereof
  • Multifunctional separation medium with tetrazole as functional group and preparing method thereof
  • Multifunctional separation medium with tetrazole as functional group and preparing method thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0035] (1) Preparation of α-aminotetrazole (R 2 Optional CH 2 CH(CH 3 ) 2 , CH(CH 3 ) 2 , CH 3 , C 6 h 5 , CH(CH 3 )CH 2 CH 3 or CH 2 C 6 h 5 )

[0036]

[0037] Synthetic route of α-aminotetrazole

[0038]Using Fmoc-protected leucine, isoleucine, valine, alanine and phenylalanine as starting materials, according to the literature method (Huo Yanmin, Lei Genhu, Zheng Xiaohui, Wei Yinmao.Synthesis and characterization of the tetrazole analogues of α-amino acids. Chinese Chemical Letters, 2006, 17(12), 1537-1539), respectively prepare Fmoc-protected α-amino tetrazole. Add Fmoc-protected α-aminotetrazole (2.5 mmol), 100 mL of 20%-50% triethylamine in methanol solution to a 250 mL round bottom flask, and stir at room temperature for 30 min. After the reaction was completed, the solvent was evaporated, and an appropriate amount of 0.5mol L was added -1 Hydrochloric acid until no solid is dissolved, extract the insoluble matter with petroleum ...

Embodiment 1

[0043] Embodiment 1: α-aminotetrazole is the preparation of the silica gel separation medium of ligand

[0044]

[0045] Preparation route of silica gel separation medium with α-aminotetrazole as ligand

[0046] Weigh 2 grams of macroporous silica gel, add 30 mL of 1:1 hydrochloric acid, ultrasonicate for 5 minutes, and then reflux in an oil bath at 100°C for 3 hours. After the reaction is complete, filter, wash with distilled water until neutral, and dry at 130°C for 4 hours. Add the silica gel into a 100mL three-neck flask, add 35mL of dry toluene, ultrasonicate for 5min, and after the temperature rises to 120°C, add 1.8mL of γ-glycidyl etheroxypropyltriethoxysilane dropwise with a dropping funnel under stirring, Reflux for 12 hours. After the reaction is completed, filter, wash with dry toluene 3 times, and wash with acetone 4 times. Vacuum dry at 50°C for later use.

[0047] Add 30mL DMF, 0.5g α-aminophenylpropane tetrazole and 0.1mL pyridine to the dry epo...

Embodiment 2

[0048] Embodiment 2: α-aminotetrazole is the preparation of the agarose (or dextran) separation medium of ligand

[0049]

[0050] α-Aminotetrazole is the preparation route of the agarose (or dextran) separation medium of the ligand

[0051] Take 10 mL of agarose (or dextran) in a 100 mL three-neck flask, add 50 mL of 5% NaOH and 5.0 mL of epichlorohydrin, and react for 12 hours. After the reaction is completed, wash with phosphate buffer (pH 7.0) to obtain agarose (or dextran) with epoxy groups bound to the surface. In epoxy agarose (or dextran), add 50mL DMF, 1.5g α-aminophenylpropane tetrazole and 1.0mL pyridine, and stir at room temperature for 24h. After the reaction is completed, wash with phosphate buffer (pH 7.0) successively to obtain tetrazolium agarose (or dextran) chromatographic separation medium.

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Abstract

The invention discloses a multi-purpose separation medium and a preparation method thereof, wherein, tetrazole is the functional group of the medium. More particularly, a separation medium with novel structure is prepared by linking the tetrazole on the surface of silica gel, dextran and agarose gel separation group. The invention has dual purposes of being applied to ion-exchange chromatography and metal chelate chromatography. When the medium is applied to the ion-exchange separation of proteins, the separation of proteins is highly selective, easy to regenerate and quick to separate and both the quality and recovery rate of the activity of the proteins are high. When the medium is applied to the metal chelate chromatography of proteins, the separation of proteins is highly selective, small in metal ion loss and quick to separate and both the quality and recovery rate of activity of the proteins are high. The invention can be applied to the quick separation and purification of genetic engineering products and plasma proteins.

Description

technical field [0001] The invention belongs to the technical field of chromatographic separation, and in particular relates to a multipurpose separation medium with tetrazole as a functional group and a preparation method thereof. Background technique [0002] Chromatographic separation technology is one of the commonly used methods for the separation of biological macromolecules in the field of bioengineering. In this method, the separation medium is the key material in the whole separation technology, and its consumption accounts for about 50% of the production cost. [0003] At present, the modes used for chromatographic separation of biological macromolecules mainly include: reversed-phase chromatography, hydrophobic interaction chromatography, ion exchange chromatography, affinity chromatography (including metal chelation chromatography) and size exclusion chromatography. Ion exchange chromatography (IEC) is the most commonly used method among them. As we all know, a...

Claims

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Application Information

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IPC IPC(8): B01J20/286C07K1/18
Inventor 卫引茂雷根虎
Owner NORTHWEST UNIV
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