Molecular imprinting method for preparing mimetic antibody and its application in 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 improved specific recognition ability, improved mechanical properties, and increased interaction sites. Effect
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[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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