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A triple-helix pH biosensor and application thereof

A biosensor, triple helix technology, used in instruments, scientific instruments, measuring devices, etc., to achieve the effects of high sensitivity, fast response, and easy operation

Active Publication Date: 2019-01-01
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the progress and development of modern environmental science, life medicine, analytical science, etc., more and more requirements are put forward for the detection of pH value, and the traditional pH detection method is facing great challenges.

Method used

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  • A triple-helix pH biosensor and application thereof
  • A triple-helix pH biosensor and application thereof
  • A triple-helix pH biosensor and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0048] In one embodiment of the present invention, the preparation method of the I chain / magnetic bead complex comprises the following steps:

[0049] Wash the carboxylated magnetic beads with a phosphate buffer solution of pH=7.0, and then activate the carboxyl group with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) solution (0.1M); Preferably, the activation conditions are: the reaction temperature is 35-40°C, and the reaction time is 20-40min;

[0050] Wash the reacted system with pH=7.0 phosphate buffer solution to remove excess EDC;

[0051] Add the aminated priming chain I chain to the above-mentioned magnetic bead dispersion for reaction to prepare the I chain / magnetic bead complex; preferably, the reaction conditions are: the reaction temperature is 35-40°C, and the reaction time is 10 ~15h.

Embodiment 1

[0053] Embodiment 1 Experimental principle

[0054] Experimental principle such as figure 1 As shown, the system consists of two hairpin DNAs (H1 and H2), a single-stranded priming DNA (I) and a capture probe (UT). The 5' end of the hairpin H1 chain and the 3' end of the H2 chain were respectively modified with the fluorescent group Alexa Fluor488. Carboxylated polystyrene magnetic beads are introduced into the system, and the I-chain of the initiator chain modified with the amino group at the 3' end can be connected to the surface of the magnetic bead through an amide reaction to obtain a I-chain / magnetic bead complex. Under acidic conditions (pH=4.0-6.0), the double-helix stems of the hairpin H1 chain and H2 chain combine with the UT chain to form triple-helix hairpin structures M1 and M2 of reverse Hoogsteen bonds. After adding the I chain / magnetic bead complex, the I chain cannot undergo the toehold-mediated chain displacement reaction. After magnetic separation, the su...

Embodiment 2

[0057] A triple-helix pH biosensor, the biosensor system includes two kinds of hairpin DNA (H1 and H2), single-stranded trigger DNA (I), capture probe (UT), the sequence of the H1 chain is 5'-TACAGCGAGGAAGGGAGAAGAGGATAGGAGTCCTCTTCTCCCTTCCTC-3 ', the sequence of the H2 chain is 5'-TCCTCTTCTCCCTTCCTCGCTGTAGAGGAAGGGAGAAGAGGACTCCTA-3', the sequence of the I chain is 5'-TCCTCTTCTCCCTTCCTCGCTGTA-3', the sequence of the UT chain is 5'-GAGGAAGGGAGAAGAGGA-3', wherein the 5' of the hairpin H1 chain The terminal and the 3' end of the H2 chain are respectively modified with the fluorescent group Alexa Fluor488, and the 3' end of the I chain is modified with an amino group. Carboxylated polystyrene magnetic beads are introduced into the system, and the 3' end of the amino group is modified. The I chain can pass through the amide The reaction attaches to the surface of the magnetic beads, resulting in I-strand / magnetic bead complexes.

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Abstract

A triple-helix pH biosensor and application thereof are disclosed. DNA modularized assembling can be achieved through changing a pH value. Accordingly, rapid and accurate signal response in a pH rangefrom 4.0 to 9.0 can be achieved based on different DNA assembling manners under different pH values. The biosensor has advantages of low cost, simple operation, rapid response, high sensitivity, andthe like. The biosensor is expected to be applied in the field of biological diagnosis based on pH-controlled DNA nanometer techniques, and has a potential application prospect in the field of drug delivery, and the like.

Description

technical field [0001] The invention relates to a triple helix pH biosensor and application thereof, belonging to the field of biosensor preparation. Background technique [0002] From chemical reactions to living organisms, whether it is a simple change or a complex reaction, pH is closely related. Monitoring pH can help people better understand changes in the microcosm. For example, the abnormal pH value of human body fluids will affect the immune system or nervous system, and even be closely related to major diseases such as cancer. Therefore, it is of great significance to accurately measure the pH value and its changes. [0003] At present, pH measurement methods mainly include pH test paper, hydrogen electrode, quinhydrone electrode, antimony electrode, Pd-H pH indicating electrode, optical fiber pH sensor, etc. The pH test paper is easy to operate, but the human error in the measurement result is large, which is not suitable for accurate measurement. Although the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6486
Inventor 毕赛齐洪杰李园芳
Owner QINGDAO UNIV
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