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Adapter-based optical chemical biosensor and method thereof for detecting target material

A biosensor and aptamer technology, applied in the field of biosensors, can solve the problems of complex modification process, cumbersome detection of target molecules, unsuitable for rapid detection of target molecules, etc., and achieve the effect of simple modification process

Inactive Publication Date: 2010-03-17
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently using aptamers as molecular recognition elements to prepare aptamer biosensors is often too complicated to modify and detect target molecules, which is not suitable for rapid detection of target molecules.

Method used

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  • Adapter-based optical chemical biosensor and method thereof for detecting target material
  • Adapter-based optical chemical biosensor and method thereof for detecting target material
  • Adapter-based optical chemical biosensor and method thereof for detecting target material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. Synthesis and amino modification of human thrombin-specific aptamer

[0038] Both the human thrombin-specific aptamer I and the human thrombin-specific aptamer II modified by amination at the 5' end were synthesized and amino-modified by Baiye Trading (Shanghai) Co., Ltd. Quality control was performed using Maldi-Tof mass spectrometry.

[0039]Aptamer I: 5'Amine-C6-TTTTTGGTTGGTGTGGTTGG

[0040] Aptamer II: 5'Amine-C6-TTTTTAGTCCGTGGTAGGGCAGGTTGGGGTGACT

[0041] All the above aptamers were dissolved in sterilized water, aliquoted and stored at -20°C. Denature at 95°C for 5 minutes before use.

[0042] 2. Preparation of human thrombin photochemical biosensor - preparation and modification of polydiacetylene nanovesicles

[0043] Dissolve PCDA in chloroform to a final concentration of 1 mmol / L. Take an appropriate amount of the solution and vacuum-dry it into a thin lipid film under nitrogen atmosphere, add deionized water to make the final concentration of the solu...

Embodiment 2

[0058] 1. Synthesis and biotin modification of human thrombin-specific aptamer

[0059] The biotinylated human thrombin-specific aptamer III and human thrombin-specific aptamer IV at the 5' end were synthesized and biotin-modified by Baiye Trading (Shanghai) Co., Ltd. Quality control was performed using Maldi-Tof mass spectrometry.

[0060] Aptamer III: 5'Biotin-C6-TTTTTGGTTGGTGTGGTTGG

[0061] Aptamer IV: 5'Biotin-C6-TTTTTAGTCCGTGGTAGGGCAGGTTGGGGTGACT

[0062] All the above aptamers were dissolved in sterilized water, aliquoted and stored at -20°C. Denature at 95°C for 5 minutes before use.

[0063] 2. Aptamer modification method based on biotin-avidin system

[0064] Dissolve PCDA and PCDA-biotin in chloroform respectively to make a final concentration of 1 mmol / L. According to the molar ratio of 1:1, an appropriate amount of solution was taken under a nitrogen atmosphere, vacuum-dried to form a thin lipid film, and deionized water was added to make the total concentrat...

Embodiment 3

[0071] 1. Synthesis and amino modification of Escherichia coli specific aptamers

[0072] The Escherichia coli-specific aptamer V modified by amination at the 5' end was synthesized by Baiye Trading (Shanghai) Co., Ltd. and completed the amino modification. Quality control was performed using Maldi-Tof mass spectrometry.

[0073] Aptamer V:

[0074] 5’Amine-C6-GCCGGCTCAGCATGACTAAGAAGGAAGTTATGTGGTGTTGGC

[0075] All the above aptamers were dissolved in sterilized water, aliquoted and stored at -20°C. Denature at 95°C for 5 minutes before use.

[0076] 2. Preparation of Escherichia coli Photochemical Biosensor-Preparation and Modification of Polydiacetylene Nanovesicles

[0077] Except that aptamer I or II in Example 1 was replaced by aptamer V, the preparation and modification of polydiacetylene nanovesicles were carried out according to the method of Example 1.

[0078] 3. Culture and counting of bacteria

[0079] Cultivate in LB liquid culture at 37°C for 16-18 hours, t...

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Abstract

The invention relates to an adapter-based optical chemical biosensor and a method thereof for detecting a target molecule, in particular to an optical chemical biosensor used for identifying a targetmolecule for example, a polydiacetylene structure organized molecular assembly of nucleotide, amino acid, polypeptide, protein, entire cell, other specific molecules and the like, by using an adapter,and a method for detecting the target molecule by using the optical chemical biosensor for the polydiacetylene structure organized molecular assembly. In the optical chemical biosensor, an adapter for a specific binding target molecule is combined on the surface of the polydiacetylene structure organized molecular assembly so that the specific target molecule can be detected / identified through the changed ultraviolet-visible light absorbency or fluorescence intensity of the polydiacetylene structure organized molecular assembly when the specific target molecule is exposed outside the corresponding adapter.

Description

technical field [0001] The invention belongs to the field of biosensors, and relates to a novel photochemical biosensor based on aptamers and a method for detecting target substances by the photochemical biosensor. Background technique [0002] Lipids containing diacetylene structures can be polymerized to form blue polydiacetylene structures under the irradiation of 254nm ultraviolet light after forming a highly ordered and compact arrangement. The color of this polydiacetylene structure will change accordingly with the temperature, solvent, pH of the environment and the change of the mechanical stress it receives. Based on this special optical property, the lipid with polydiacetylene structure becomes an ideal material for preparing signal transduction elements of biosensors. [0003] In 1993, D.H.Charych and others published an article in Science about the ability to identify influenza viruses by using functionalized polydiacetylene color-changing films, which aroused wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/64G01N33/52G01N33/569C12Q1/68C12Q1/37C12Q1/04C12R1/19
Inventor 吕建新陈勇吴文鹤
Owner WENZHOU MEDICAL UNIV
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