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Method for preparing agglutinin high-performance affinity chromatography material by taking silica gel as substrate

A technology of lectin and silica gel, which is applied in the field of preparation of lectin-affinity materials, can solve problems such as poor maintenance of lectin activity, lengthy and complicated preparation routes, etc., and achieve small loss, simple operation and stable performance Effect

Active Publication Date: 2016-06-01
中科榆林能源技术运营有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the problems that the preparation route of lectin affinity chromatography material is lengthy and complicated, and the activity of lectin cannot be well maintained, the present invention provides a method for efficiently preparing lectin affinity material, which mainly involves the spacer arm of the affinity material Synthesis, Synthetic Process of Lectin Affinity Materials

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Take 5g of silica gel, put it in a 100mL glass round bottom flask, add 50mL of 6M hydrochloric acid, heat to reflux for 1 hour, wash with water until neutral, and dry at 150°C for 5 hours.

[0025] Under nitrogen protection, take 1.66g of aminopropyltrimethoxysilane reagent, dissolve it in 20mL of anhydrous THF solution, stir it electromagnetically, add 1.027g of succinic anhydride, adjust the pH value to 7 with triethylamine, stir at room temperature, and perform liquid chromatography Carboxylated silane reagents were detected.

[0026] Under nitrogen protection, the dried silica gel (2.00g) was dissolved in 30mL of anhydrous toluene, the above-mentioned silane reagent containing carboxyl functional groups was added, and pyridine (1mmol, 80uL) was added dropwise, heated to reflux, reacted for 12 hours, and stopped the reaction. Cool and suction filter, wash with anhydrous toluene, tetrahydrofuran, methanol, water, and methanol successively, and dry and solidify at 80°C...

Embodiment 2

[0028] Example 2: Take 5g of silica gel, place it in a 100mL glass round-bottom flask, add 50mL of 6M hydrochloric acid, heat to reflux for 4 hours, wash with water until neutral, and dry at 150°C for 5 hours;

[0029] Protected under a nitrogen atmosphere, take 1.66 g of aminopropyltriethoxysilane reagent, dissolve it in 10 mL of anhydrous THF solution, stir it electromagnetically, add 1.027 g of glutaric anhydride, adjust the pH to 6 with triethylamine, and stir at room temperature. The silane reagent containing carboxyl group at the end was obtained by liquid chromatography detection. Under nitrogen protection, dissolve the dried silica gel (2.00g) in 20mL of anhydrous toluene, add a silane reagent containing a carboxyl functional group, and dropwise add pyridine (1mmol, 80μL), heat to reflux, react for 48 hours, stop the reaction, and cool Suction filtration, washing with anhydrous toluene, tetrahydrofuran, methanol, water, methanol successively, drying and curing at 80°C ...

Embodiment 3

[0031] Example 3: under nitrogen protection, the dried silica gel (2.00g) was dissolved in anhydrous toluene, glycidyloxypropyltrimethoxysilane (0.7ml) was added, and triethylamine was added dropwise to adjust the pH value 8, heated to reflux, reacted for 48 hours, stopped the reaction, cooled and filtered, washed with anhydrous toluene, tetrahydrofuran, methanol, water, methanol in turn, dried and solidified at 80°C for 12 hours, and obtained silica gel immobilized with epoxy functional groups at the end Mutually.

[0032] Under nitrogen atmosphere protection, weigh 1.0 g of the above-mentioned dry silica gel and dissolve it in 20 mL (pH 7.2) MES buffer solution; take 100 mg soybean agglutinin (SBA) and add it to 0.1M N-acetyllactosamine MES buffer saline solution, stir well, Add it to the above silica gel, and stir at 4°C for 12 hours; after the reaction is completed, add ethanolamine with a final volume concentration of 2% to block the activating group, stir for 1 hour, cen...

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PUM

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Abstract

The invention relates to the technical field of an affinity material, silica gel is taken as a substrate, a preparation method comprises synthesis of a spacer arm-modified silica gel containing bifunctional groups, and is used for preparing an agglutinin affinity material containing high bonding amount. A structure of the agglutinin affinity material is characterized in that the silica gel is taken as the substrate, the spacer arm end contains function groups of epoxy, hydroxyl, carboxyl, isocyano, and aldehyde group, and a ligand is agglutinin. The prepared agglutinin affinity material has the advantages of simple process, mild condition, low cost, high bonding amount, stable agglutinin property, and good agglutinin activity, and can be widely used for separating and gathering glycoprotein and glycopeptides.

Description

technical field [0001] The invention relates to the preparation technology of affinity materials, mainly the preparation method of lectin affinity materials, and relates to the synthesis of spacer arms and the preparation of lectin affinity materials. Background technique [0002] Glycosylation of proteins is one of the most important modifications in protein post-translational modifications. More than 50% of proteins in organisms are glycosylated. Glycoproteins are involved in cellular immunity, signal transduction, protein translation regulation, cancer and other physiological The process plays an important role in living organisms (Syntheticglycopeptides and glycoproteins as tools for biology, Chem. Soc. Rev., 2005, 34, 58-68; Cancerglycan biomarkers and their detection–past, present and future, Anal. Methods, 2014, 6, 3918-3936). 正因为疾病中存在着异常的糖基化现象(ElucidationofN-GlycositesWithinHumanPlasmaGlycoproteinsforCancerBiomarkerDiscovery,MassSpectrometryofGlycoproteins,2013,951,30...

Claims

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Application Information

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IPC IPC(8): B01J20/281B01J20/24B01J20/30
Inventor 刘玉洁梁鑫淼于龙付冬梅肖远胜
Owner 中科榆林能源技术运营有限责任公司
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