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Deoxyribonucleic acid (DNA)-modified gold nanoparticle colorimetric sensor and preparation method and use thereof

A colorimetric sensor and gold nanotechnology, applied in the field of bioanalysis, achieves high selectivity, low energy consumption, and simple preparation

Active Publication Date: 2014-07-16
ANHUI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the inherent defects of these methods, such as requiring expensive and complex instruments, limit their development as an important means of real-time detection of FR

Method used

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  • Deoxyribonucleic acid (DNA)-modified gold nanoparticle colorimetric sensor and preparation method and use thereof
  • Deoxyribonucleic acid (DNA)-modified gold nanoparticle colorimetric sensor and preparation method and use thereof
  • Deoxyribonucleic acid (DNA)-modified gold nanoparticle colorimetric sensor and preparation method and use thereof

Examples

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Embodiment 1

[0035] A DNA-modified gold nanocolorimetric sensor for the colorimetric detection of folate receptors based on tail protection. A method for preparing a gold nanoparticle colorimetric sensor, specifically comprising:

[0036] a. Dissolve the purchased DNA sequences (thiol: S1, FA-DNA: S2) in 0.05M Tris-HCl (pH7.4) buffer solution, and store them at 4°C for future use.

[0037] b. Using the succinimide coupling method (EDC-NHS) to bind FA to S2 with a modified amino group at the 3' end. 0.5mL of 20μM S2 was mixed with 0.5mL of 100mM Tris-HCl buffer solution (pH7.4, containing 10mM FA, 1mM EDC, 5mM Sulfo-NHS) and incubated at 37°C in the dark for 3h. The FA-S2 solution was dialyzed in PBS buffer solution (pH 7.4) to remove excess FA (molecular weight cutoff: 1000 Da). Then FR with standard serial concentration was added to the above-mentioned dialyzed solution, and cultured at 37° C. in the dark for another 2 h. Finally, 400U / mL Exo I was added to the above mixed solution, an...

Embodiment 2

[0040] A DNA-modified gold nanocolorimetric sensor for the colorimetric detection of folate receptors based on tail protection. The preparation and application steps of the colorimetric sensor based on S1-AuNP and S2 / FA-FR are as follows:

[0041] a. Dissolve the purchased DNA sequences (thiol: S1, FA-DNA: S2) in Tris-HCl (pH7.4) buffer solution, and store them at 4°C for future use.

[0042] b. Using the succinimide coupling method (EDC-NHS) to bind FA to S2 with a modified amino group at the 3' end. A certain concentration of S2 and Tris-HCl buffer solution (pH7.4, containing 10mM FA, 1mM EDC, 5mM Sulfo-NHS) were mixed evenly in a certain ratio, and incubated in the dark at 37°C for several hours. The FA-S2 solution was dialyzed against PBS buffer solution (pH 7.4) to separate excess FA (molecular weight cutoff: 1000 Da). Then add standard serial concentration of FR into the above dialyzed solution, and incubate for a period of time in the dark at 37°C. Finally, 400U / mL E...

Embodiment 3

[0045] Preparation of S1-Au NPs: Using a chemical reduction method, first synthesize a dispersion solution of Au NPs with uniform size (14nm), and then modify DNA-S1 with sulfhydryl groups to the surface of the particles.

[0046] The preparation and application steps of the colorimetric sensor based on S1-Au NPs and S2 / FA-FR are as follows:

[0047] a. Dissolve the purchased DNA sequences (thiol: S1, FA-DNA: S2) in 0.05M Tris-HCl (pH7.4) buffer solution, and store them at 4°C for future use.

[0048]b. Using the succinimide coupling method (EDC-NHS) to bind FA to S2 with a modified amino group at the 3' end. 0.5mL of 20μM S2 was mixed with 0.5mL of 100mM Tris-HCl buffer solution (pH7.4, containing 10mM FA, 1mM EDC, 5mM Sulfo-NHS) and incubated at 37°C in the dark for 3 hours. The FA-S2 solution was dialyzed against PBS buffer solution (pH 7.4) to separate excess FA (molecular weight cutoff: 1000 Da). Then FR with standard serial concentration was added to the above-mentione...

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Abstract

The invention relates to a deoxyribonucleic acid (DNA)-modified gold nanoparticle colorimetric sensor and a preparation method and use of the DNA modified gold nanoparticle colorimetric sensor. The gold nanoparticle is synthesized by using a chemical reduction method; DNA (S1) of modifying sulfydryl is modified on the particle surface to prepare S1-Au NPs by using a gold sulfur bond; FR can be specifically combined with a folic acid (FA) to form FR-FA, so that the DNA (FA-DNA) modified by FA is not hydrolyzed by nucleotide circumscribed enzyme-I (Exo I), so as to protect the FA-DNA. The S1-Au NPs colorimetric sensor is built by utilizing the property and the complementary pairing principle of basic groups between DNA sequences, and direct selective detection of FR is achieved. The colorimetric sensor disclosed by the invention is simple to prepare, low in energy consumption, high in selectivity, fast and visual, and applicable to real-time detection of FR, and can be expected to be applied to clinical diagnosis.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and in particular relates to the preparation of a gold colloid solution colorimetric sensor and its detection of folic acid receptors. Background technique [0002] Folate receptor (FR) plays a vital role in the process of physiology, pharmacology and cell pathology, so it is of great significance to directly and selectively detect the expression of FR in malignant tumors. In the past 20 years, fluorescence resonance energy transfer (FRET), fluorescence anisotropy, capillary electrophoresis (CE), surface plasmon resonance (SPR), etc. are the most commonly used methods for detecting FR. However, the inherent defects of these methods, such as requiring expensive and complicated instruments, limit their development as an important means for real-time detection of FR. Contents of the invention [0003] The object of the present invention is to provide a DNA-modified gold nanometer colo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 王广凤朱艳红陈玲
Owner ANHUI NORMAL UNIV
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