A preparation method of molecularly imprinted composite material for enrichment and separation of glycoproteins

A composite material, molecular imprinting technology, applied in the fields of materials science and engineering and bioseparation engineering, can solve problems such as unfavorable protein binding and loading, limited protein affinity, etc. , fast binding and elution

Active Publication Date: 2018-07-31
沛县度创科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the surface of graphene is inert and has limited affinity with proteins, which is not conducive to the binding and loading of proteins on its surface.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] (1) Synthesis of borate-based functionalized graphene

[0019] 40 mg of graphene oxide was added to 200 mL of deionized water, and the solution was uniformly dispersed by ultrasonic for 30 min to obtain an aqueous solution of graphene oxide. Add 240mg of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC·HCl) and 240mg of N-hydroxysuccinimide (NHS) to the above solution, and stir for 20 min. , Add 400 mg of 3-aminophenylboronic acid monohydrate (APBA), stir at room temperature for 18 hours, suction filter, wash with ethanol and water 3 times, suction filter, and vacuum dry to obtain boric acid-based functionalized graphene oxide.

[0020] (2) Preparation of molecularly imprinted composite materials

[0021] Take 40 mg of the boronic acid-based functionalized graphene obtained in the previous step, disperse it in 40 mL of PBS (pH=6.5) buffer solution containing 200 mg of template glycoprotein, and react for 6 hours at room temperature to fully assemble the glycopr...

Embodiment 2

[0023] (1) Synthesis of borate-based functionalized graphene

[0024] 400 mg of graphene oxide was added to 200 mL of deionized water, and the solution was uniformly dispersed by sonicating for 40 minutes to obtain a graphene oxide aqueous solution. Add 1.2g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) and 1.2g of N-hydroxysuccinimide (NHS) to the above solution, and stir After 30 minutes, add 2.4 g of 3-aminophenylboronic acid monohydrate (APBA), stir at room temperature for 24 hours, and then suction filter, wash with ethanol and water, suction filter, and vacuum dry to obtain boric acid-based functionalized graphene oxide;

[0025] (2) Preparation of molecularly imprinted composite materials

[0026] Take 350 mg of the boronic acid-based functionalized graphene obtained in the previous step, disperse it in 40 mL of PBS (pH = 7.4) buffer solution containing 180 mg of template glycoprotein, and react for 10 hours at room temperature to fully assemble the g...

Embodiment 3

[0028] (1) Synthesis of borate-based functionalized graphene

[0029] 400 mg of graphene oxide was added to 100 mL of deionized water, and the solution was uniformly dispersed by ultrasonic for 80 min to obtain an aqueous solution of graphene oxide. Add 800 mg of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC ·HCl) and 800 mg of N-hydroxysuccinimide (NHS) to the above solution, and stir for 40 min. , Add 1.2g of 3-aminophenylboronic acid monohydrate (APBA), stir at room temperature for 30h, suction filter, wash with ethanol and water, suction filter, and vacuum dry to obtain boric acid-based functionalized graphene oxide;

[0030] (2) Preparation of molecularly imprinted composite materials

[0031] Take 400 mg of the boronic acid-based functionalized graphene obtained in the previous step, disperse it in 50 mL of PBS (pH = 8.5) buffer solution containing 400 mg of template glycoprotein, and react for 10 hours at room temperature to fully assemble the glycoprotein i...

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PUM

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Abstract

The invention discloses a preparation method of a molecularly imprinted composite material for enriching and separating glycoproteins, belonging to the fields of material science and engineering and biological separation engineering. In this method, the surface of graphene oxide is firstly functionalized with boric acid, the template glycoprotein is fixed on the surface of graphene oxide through boron affinity, and the surface imprinting technology is used to form a protein imprinted layer on the surface of graphene oxide, and the template is eluted to obtain molecular imprinting. composite material. The molecularly imprinted composite material prepared by this method has a very good ability to specifically recognize the template glycoprotein, and also has the advantages of high binding capacity, strong anti-interference ability, fast binding and elution speed, and can be used for complex practical applications. The low-concentration glycoproteins in the sample are enriched, separated and detected.

Description

Technical field: [0001] The invention discloses a preparation method of a molecularly imprinted composite material for enriching and separating glycoproteins, and belongs to the fields of material science and engineering and biological separation engineering. Background technique: [0002] Glycoprotein is a relatively common type of protein in organisms, which has important biological functions and clinical significance. Glycoproteins play important roles in physiological processes, such as molecular recognition, immune response and developmental regulation, and many diseases are related to the expression of glycoproteins. Therefore, many glycoproteins have been used as disease biomarkers for clinical diagnosis. For early diagnosis of major diseases, such as infection, tumor, cardiovascular disease, liver disease, kidney disease and diabetes, etc. Common disease markers such as embryonic antigen (CEA), alpha-fetoprotein (AFP) and prostate specific antigen (PSA) are all glycoprot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F220/56C08F222/38C08K9/04C08K3/04C08J9/26B01J20/26B01J20/30
Inventor 罗静丛姣姣郑媛高雅涵刘晓亚
Owner 沛县度创科技发展有限公司
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