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Protein antigen determinant molecularly imprinted material as well as preparation and application thereof

An epitope and molecular imprinting technology, which is applied in the direction of material inspection products, other chemical processes, chemical instruments and methods, etc., can solve the problems of low template utilization rate, difficult removal of templates, and limitations on the application of epitope imprinting methods, and achieve Stable conformation, simple method, guaranteed versatility and universal applicability

Inactive Publication Date: 2015-09-30
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the template utilization rate is extremely low when free peptides are used as templates, the templates need to be immobilized; however, the template immobilization methods reported previously have the disadvantage that templates are difficult to remove, which limits the application of epitope imprinting methods

Method used

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  • Protein antigen determinant molecularly imprinted material as well as preparation and application thereof
  • Protein antigen determinant molecularly imprinted material as well as preparation and application thereof
  • Protein antigen determinant molecularly imprinted material as well as preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] (1) Preparation of human serum albumin epitope imprinted microsphere material

[0029] Magnetic nanoparticles coated with silicon layer were prepared as matrix material by solvothermal method. Take 50 mg of matrix material, add 1ml of 0.5mg / ml human serum albumin template peptide (AASQAALGLHHHHHH) solution (Gill Biochemical Shanghai Co., Ltd.), incubate at 25°C to immobilize the template on the matrix surface (the mass ratio of the template to the matrix material is 10:1000). After washing the unfixed template peptides, resuspend the matrix in 25ml of water, add 15mg of dopamine (mass ratio of monomer to matrix material is 0.3:1), pre-assemble under mechanical stirring at 30°C for 30min, add 0.5ml of 1M tris-HCl buffer (pH8.0), self-polymerized for 5h under mechanical stirring, and cleaned unpolymerized dopamine monomers with the assistance of a magnet. Subsequently, the magnetic particles were repeatedly washed with 200 mM EDTA-Na to make Ni 2+ After elution, the te...

Embodiment 2

[0038] It is 40:1000 to carry out template immobilization by template and matrix material mass ratio, other can obtain MIP2 and NIP2 with embodiment 1, its adsorption capacity to human serum albumin epitope is as follows Figure 4 shown.

Embodiment 3

[0040] It is 2.5:1000 to carry out template immobilization by template and matrix material mass ratio, other can obtain MIP3 and NIP3 with embodiment 1, its adsorption capacity to human serum albumin epitope is as follows Figure 5 shown.

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Abstract

The invention relates to an antigen determinant molecularly imprinted material capable of carrying out selective recognition on a target protein and a preparation process of the antigen determinant molecularly imprinted material. The molecularly imprinted material is prepared by the following steps: by utilizing a section of specific peptide fragment corresponding to the target protein as a template molecule, fixing the specific peptide fragment on a matrix material by employing a histidine tag as a connecting arm; and polymerizing on the matrix material through a functional monomer. The imprinted material is applied to selective recognition of a standard protein.

Description

technical field [0001] The invention belongs to polymer material preparation technology and its application in protein recognition, specifically a novel protein epitope molecular imprinting material and its preparation and application. Background technique [0002] Molecular imprinting technology is a technology that uses artificial synthesis to prepare materials for specific recognition of target molecules. The earliest related report was published in Nature by Klaus Mosbach et al. in 1993 on the use of non-covalent bonds to synthesize theophylline molecularly imprinted polymers. Since then, molecularly imprinted materials have rapidly entered the world as a synthetic high-selectivity material. The scientist's perspective. At present, great progress has been made in the imprinting of small organic molecules and metal ions, and the experimental methods and evaluation standards are gradually becoming mature, and have begun to be applied in clinical analysis, catalytic synthe...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/30C08J9/26C08J7/16C08J7/12G01N33/68C04B41/48
Inventor 张丽华李森武杨开广李沁然张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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