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Novel molecular imprinting method for preparing simulation antibody and application of novel molecular imprinting method on bacterial detection

A technology of molecular imprinting and imitating antibodies, which is applied in the measurement of color/spectral characteristics, etc., can solve the problems of poor specific recognition ability and low sensitivity, and achieve the effect of improving specific recognition ability, improving mechanical properties, and widely promoting application value

Inactive Publication Date: 2016-09-28
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the problems of poor specific recognition ability and low sensitivity of the existing surface molecular imprinting technology, and to provide a molecular imprinting method for preparing a mimetic antibody with strong specific recognition ability, high sensitivity, low cost, and easy operation. New method, and the application effect on bacterial detection is superior

Method used

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  • Novel molecular imprinting method for preparing simulation antibody and application of novel molecular imprinting method on bacterial detection
  • Novel molecular imprinting method for preparing simulation antibody and application of novel molecular imprinting method on bacterial detection

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preparation example Construction

[0017] In the new method of molecular imprinting for preparing mimetic antibodies described in the present invention, at least two polymers with different characteristics and different functional groups are blended into a solution in a certain proportion, and then the inactivated bacterial solution is mixed with the blended solution , and then the mixed solution was spin-coated on the porous gold film by the spin-coating method, and the mixed solution spin-coated on the porous gold film was slowly dried at room temperature to form a blended film compounded with bacteria, and in the process of film formation In , the induced assembly of the polymer and the bacterial surface occurs (eg figure 1 Shown), different functional groups and the surface of the bacteria form a site that simulates the mutual recognition of antigen-antibody, and then the bacterial template is eluted from the blended membrane compounded with bacteria, and then slowly dried at room temperature to form a molec...

Embodiment 1

[0021] Put polyvinyl alcohol (PVA) in water, heat up to 80°C, and stir until it is completely dissolved; put gelatin in water, heat up to 40°C, and stir until it is completely dissolved; then mix the prepared PVA solution with gelatin The solution is mixed in a mass ratio of 9:1, and then the inactivated Escherichia coli solution is mixed with the blended solution of PVA / gelatin, and the mixed solution is spin-coated on the porous gold film by the spin coating method, and the coating thickness is 1μm, slowly dry at room temperature to form a film. The gelatin / polyvinyl alcohol film (MIP film) compounded with E. coli prepared by spin coating was first pretreated with lysozyme (10mg / ml) for 2 hours at 4°C to hydrolyze and destroy the cell wall of E. coli. The MIP membrane was then treated with 10% Triton X for 80 minutes to remove the strong interaction between the polysaccharides on the cell wall and the polymer surface, followed by washing the MIP membrane with a large amount ...

Embodiment 2

[0023] Place the agarose in water, raise the temperature to above 90°C, and stir at a constant temperature until it is completely dissolved; place the chitosan in water, adjust the pH value of the water to a weak acid, and stir at a constant temperature until it is completely dissolved; then the prepared agarose solution Mix with chitosan solution at a mass ratio of 1:1, then mix inactivated Escherichia coli solution with agarose / chitosan blend solution, and spin-coat the mixed solution on porous gold by spin coating method. On the film, the coating thickness is 1 μm, and the film is slowly dried at room temperature. The agarose / chitosan membrane (MIP membrane) compounded with Escherichia coli prepared by spin coating was first pretreated with lysozyme (10mg / ml) for 2 hours at 4°C to hydrolyze and destroy the cell wall of Escherichia coli , and then treated the MIP membrane with 10% Triton X for 80 minutes to remove the strong interaction between the polysaccharides on the cel...

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Abstract

The invention discloses a novel molecular imprinting method for preparing a simulation antibody and application of the novel molecular imprinting method on bacterial detection. According to the novel molecular imprinting method, a molecular imprinting film is prepared by blending two polymers with different properties and functional groups in proportion; uncross-linked blended polymers are particularly applied to the molecular imprinting technique of the simulation antibody, and the mutual action sites between the imprinting film and bacteria are increased by virtue of mutual action of simulation antigens-antibodies between the different functional groups of the polymers and the surfaces of the bacteria, so that the specific recognition capacity of the imprinting film is improved. According to the novel molecular imprinting method, the problems of poor specific recognition capacity and low sensitivity of a surface molecular imprinting technique can be solved, and a bacteria detection method having the characteristics of strong specific recognition capacity, high sensitivity, low cost, simplicity and feasibility can be provided.

Description

technical field [0001] The invention relates to a new molecular imprinting method for preparing highly simulated mimetic antibodies based on an interface-induced assembly method and its application in bacterial detection. Background technique [0002] Molecular Imprinting Technique (MIT) is a new experimental preparation technology for artificially synthesizing polymers coupled with template molecules. Its basic idea comes from people's understanding of the specific selection of antigen-antibody and enzyme-substrate , when the template molecules are in contact with the polymer monomers, multiple interaction points are formed, and this effect will be memorized during the polymerization process. Cavities with multiple sites of action, such cavities will have selective recognition properties for template molecules and their analogs. The polymers prepared by MIT that completely match the template molecules in terms of spatial structure and binding sites are also called molecula...

Claims

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Application Information

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IPC IPC(8): G01N21/33C08L29/04C08L89/00C08L5/12C08L5/08C08J5/18
CPCG01N21/33C08J5/18C08L5/08C08L5/12C08L29/04C08J2405/12C08J2405/08C08J2489/00C08J2305/12C08J2329/04C08J2305/08
Inventor 孙康周建华冯罕博
Owner SUN YAT SEN UNIV
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