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Microbiological bio-imprinting membrane and preparation method thereof

A technology of bio-imprinting and microorganisms, applied in the direction of layered products, etc., can solve the problems of too large target substances, difficulties in immobilization, and difficulties in microbial bio-imprinting, and achieve the effects of mild deposition conditions, increased biocompatibility, and easy operation

Inactive Publication Date: 2012-06-27
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are still relatively few imprinting techniques for micron-scale substances such as microorganisms, mainly because the target substance is too large, and it is difficult to move and fix.
Moreover, the commonly used methods of molecular imprinting usually involve organic solvents. The organic solvent itself will cause the cell wall of microorganisms to dissolve, cross-link and other reactions. These problems have brought many difficulties to the bioimprinting of microorganisms.

Method used

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  • Microbiological bio-imprinting membrane and preparation method thereof
  • Microbiological bio-imprinting membrane and preparation method thereof
  • Microbiological bio-imprinting membrane and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Preparation steps of graphene:

[0026] 1g natural graphite, 1g NaNO 3 It was mixed with 46 mL of concentrated sulfuric acid (98%) and reacted in an ice-water bath for 4-5 h. Then 6g KMnO 4 Added to the mixture solution while raising the temperature to 35°C and maintaining the reaction temperature, after the reaction was carried out for 2-3h, 90-95mL of H was added 2 O and the temperature was raised to 98 °C, after the reaction was carried out for two hours, 200 mL of warm water, 20 mL of 30% H 2 O 2 , and continue the reaction at room temperature for 1-2h. Always keep stirring during the whole reaction process. The prepared product was filtered, washed with distilled water and dried in an oven at 60° C. for 24-48 h to obtain a yellow solid. After the reaction product was ground into pieces, 0.1 g was weighed and added to 100 mL of H 2 Sonicate in O for 1-2h to obtain a homogeneous mixed solution, add 2-3mL N 2 H 4 Then, it is divided into a reactor and reacted ...

Embodiment 2

[0028] Preparation steps for microbial bioblotting films (see figure 1 ):

[0029] 1) Cutting and cleaning of ITO electrodes. Cut the ITO electrode into glass pieces of uniform size (1 × 4 cm) with a glass knife, then add absolute ethanol and ultrapure water to the beaker in turn, and ultrasonically treat them for 30 minutes respectively. After taking them out, measure the conductive surface with a multimeter. dry, spare;

[0030] 2) Deposition of graphene / chitosan composite membrane carrier. The graphene prepared in Example 1 was centrifugally cleaned and added to a chitosan solution with a mass fraction of 0.5% to obtain a graphene-containing concentration of 0.3 mg mL -1 Graphene / chitosan mixture. Using the potentiostatic deposition method, a three-electrode system was used, the platinum wire electrode was used as the counter electrode, the Ag / AgCl electrode was used as the reference electrode, the ITO electrode in step 1) was used as the working electrode, and the depo...

Embodiment 3

[0036] Selectivity Evaluation of Microbial Bioblot Films:

[0037] The selectivity of the microbial bioimprinted film obtained in Example 2 was evaluated by electrochemical impedance method. A three-electrode system was adopted, the platinum wire electrode was used as the counter electrode, the Ag / AgCl electrode was used as the reference electrode, and the ITO electrode with the microbial bioimprinting film prepared in Example 2) was used as the working electrode. 6 ] 4- / 3- The 0.2M PBS (pH 7.4) buffer was used as the electrolyte. The experimental parameters of electrochemical impedance spectroscopy were set to be measured at open circuit potential, the amplitude of the sine wave was 10mV, and the frequency range was from 0.1Hz to 100kHZ. Rod-shaped bacteria and spherical bacteria of the same concentration were respectively added dropwise to the ITO electrode modified with the recognition site of microbial bioprinting obtained in Example 2. By measuring the electrochemical ...

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Abstract

The invention relates to the field of molecular imprinting and specifically to a microbiological bio-imprinting membrane and a preparation method thereof. According to the invention, a chitosan / graphene composite membrane, a microbial template and a chitosan membrane are sequentially deposited on a conductive substrate so as to prepare the microbiological bio-imprinting membrane. The preparation method for the microbiological bio-imprinting membrane has the advantages of simple preparation process and mild preparation conditions; the prepared microbiological bio-imprinting membrane can be used in the fields of screening, separation and detection of microbes, etc.

Description

technical field [0001] The invention relates to the field of molecular imprinting, in particular to a microbial imprinting film and its preparation. Background technique [0002] Molecular imprinting technology is a technology for synthesizing artificial antibodies. Its main principle is to select functional monomers that can form covalent or non-covalent bonds with template molecules, form complexes with template molecules, and then copolymerize through cross-linking agents. Cross-linked polymers are formed. After the template molecule is eluted, holes are left in the polymer that match the shape, size and surface groups of the template molecule. The target molecules are recognized through these holes, that is, biological recognition sites. The use of molecular imprinting technology to make artificial antibodies as biological recognition elements has become a popular direction of biosensor research in recent years. Molecular imprinting technology can be applied in cataly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B32B9/00C08L5/08C08K3/04C08J9/26
Inventor 戚鹏张盾万逸
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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