Molecularly imprinted polymer specially combined with specified glycoprotein and preparation method and application of molecularly imprinted polymer

A technology of molecular imprinting and glycoprotein, which is applied in the photoplate making process of patterned surface, biological testing, material inspection products, etc., can solve the problems of many steps, poor anti-interference ability, long cycle, etc., and achieve simple preparation and anti-interference Strong ability and high specific effect

Inactive Publication Date: 2012-06-27
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these current methods still have shortcomings such as many preparation steps, long cycle time and poor anti-interference ability. More importantly, there is no universally applicable method for molecular imprinting of glycoproteins.

Method used

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  • Molecularly imprinted polymer specially combined with specified glycoprotein and preparation method and application of molecularly imprinted polymer
  • Molecularly imprinted polymer specially combined with specified glycoprotein and preparation method and application of molecularly imprinted polymer
  • Molecularly imprinted polymer specially combined with specified glycoprotein and preparation method and application of molecularly imprinted polymer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Preparation of horseradish peroxidase (HRP) imprinted polymer

[0035] The preparation process of HRP molecularly imprinted polymer is as follows:

[0036] 1. Prepare the prepolymer solution, dissolve 0.005g p-vinylphenylboronic acid and 0.001g benzoin dimethyl ether in 400μl polyethylene glycol 200 (PEG200, average molecular weight 200), 100μl polyethylene glycol diacrylate ( PEGMA), phosphate buffer solution to adjust the pH value to 8.5, add 5 μl of 10 mg / ml HRP solution, and vortex for 2 minutes (the mass ratio of glycoprotein HRP to functional monomer p-vinylphenylboronic acid is 1: 100).

[0037] 2. Drop 100μl of pre-polymerization solution on the glass substrate derivatized with double bonds, attach a mask plate containing a specific structure, and expose to UV (365nm) for 40s. Remove the mask, soak the polymer in a mixed solution of ethanol / water (40% ethanol by volume) and vibrate for 30 minutes, then soak the polymer in acetonitrile / 10M phosphoric...

Embodiment 2

[0040] Example 2: Recognition of HRP by HRP imprinted polymers and comparison with non-imprinted polymers

[0041] The HRP imprinted polymer and non-imprinted polymer were used to identify HRP respectively, and 3,3',5,5'-tetramethylbenzidine (TMB) chromogenic method was used for naked eye detection:

[0042] 1. Prepare TMB chromogenic solution, sodium acetate 0.136g, citric acid 0.016g, 30% hydrogen peroxide 3μl, TMB 1.5mg, distilled water to 5ml.

[0043] 2. In non-imprinted polymers ( image 3 Add 5 μl of water (pH 6.0), HRP aqueous solution (10 μg / ml, pH 6.0) and HRP phosphate solution (10 μg / ml, pH 8.5) dropwise on columns A, B, and C of A), and after 2 minutes add acetonitrile Rinse the non-imprinted polymer with a mixed solution of water / water (volume ratio 30:70) for 1 minute, dry the polymer, add 5 μl of TMB chromogenic solution dropwise to each point, and react in the dark for 10 minutes, and the polymerization after color development Objects recorded with a digital...

Embodiment 3

[0046] Example 3 Anti-interference ability of molecularly imprinted polymer

[0047] Using the same preparation steps as in Example 1, the pH value of the phosphate buffer solution in the formula was adjusted to 8 to prepare the HRP imprinted polymer. The recognition of HRP by HRP imprinted polymers in complex systems was observed by naked eyes using TMB chromogenic method:

[0048] 1. Prepare a complex solution: 1) dissolve 50 μg HRP, 1 mg fructose, 1 mg ovalbumin, 1 mg human hemoglobin, and 1 mg bovine serum albumin in 1 ml human serum; 2) dissolve 10 μg HRP, 1 mg fructose, 1 mg ovalbumin, 1 mg human Hemoglobin, 1 mg bovine serum albumin were dissolved in 1 ml human serum; 3) 5 μg HRP, 1 mg fructose, 1 mg ovalbumin, 1 mg human hemoglobin, 1 mg bovine serum albumin were dissolved in 1 ml human serum.

[0049] 2. Imprinted polymers in HRP ( Figure 4 Add 5 μl of water, serum, solution 1), solution 2), and solution 3) dropwise on columns 1, 2, 3, 4, and 5 of A), and after 2 m...

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Abstract

The invention discloses a molecularly imprinted polymer specially combined with a specified glycoprotein. Substituted phenylboronic acid containing double bonds is used as a functional monomer. Under the alkali condition, the specified glycoprotein serving as a template (imprinting molecule) and the substituted phenylboronic acid (functional monomer) containing the double bonds form a complex; the complex is mixed with a cross linker, an initiator and a pore forming agent; copolymerization reaction between the functional monomer and the cross linker and in the cross linker is initiated under the irradiation of ultraviolet light to form a polymer; and the template modules (specified glycoprotein) in the polymer are removed by extraction of an acidic solution, and thus the molecularly imprinted polymer containing a phenylboronic acid site reversibly combined with the glycosyl of the glycoprotein and a cavity complementary with the shape of the template modules is obtained. The molecularly imprinted polymer specially combined with the specified glycoprotein has the advantages of high specificity, strong anti-interference capacity, wide practical sample range, good repeatability, simplicity in preparation and the like, and can be directly used for detecting antigens to be detected in a complex biological system of serum, saliva and the like. The invention discloses a preparation method for the molecularly imprinted polymer.

Description

technical field [0001] The invention relates to a molecularly imprinted polymer specifically binding to glycoprotein, its preparation method and application in detecting antigen glycoprotein. Background technique [0002] Glycoproteins are an important class of proteins. In mammals, more than half of the total protein is glycoprotein. Glycoproteins play important roles in physiological processes such as molecular recognition, intra / intercellular signaling, immune response, fertilization, and developmental regulation, among others. Glycoproteins are of great significance in the early diagnosis of major diseases. Most of the disease markers approved by the US Food and Drug Administration (FDA) are glycoproteins, such as alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA) and prostate specific antigen (PSA). Furthermore, glycoproteins such as erythropoietin (EPO) and human chorionic gonadotropin (hCG) are banned substances in sports, and the rapid and sensitive detection...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F230/06C08F222/14C08F222/38C08F2/44C08F2/48C08J9/28G03F7/00G01N33/68
Inventor 刘震李澧
Owner NANJING UNIV
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