Method for detecting glutathione

A glutathione and probe technology, applied in the field of GSH detection, to achieve excellent selectivity and specificity, high selectivity detection, and sensitive fluorescence signal effects

Active Publication Date: 2017-06-30
合肥龙之韵医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using gold nanocages as nanocarriers and using biomolecules containing disulfide bonds to construct fluorescent sensors based on controlled release to detect GSH has not been reported in the literature.

Method used

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  • Method for detecting glutathione

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Embodiment 1

[0020] A method for preparing the GSH fluorescent sensor based on the controllable release technology proposed by the present invention, comprising the steps of:

[0021] (1) Mix 20 μL of washed mercapto magnetic beads with 400 μL of gold nanocage solution evenly, shake at a constant temperature of 37°C for 12 hours, magnetically separate, wash, and add 200 μL of polydiallyldimethyl to the prepared gold nanocage complex Ammonium chloride solution (11.664mg / mL), shake at 37°C for 12h, magnetically separate and wash;

[0022] (2) Add 100 μL rhodamine B solution (1.0×10 -5 mol / L), 10 μL of GSH recognition probe solution (1.0×10 -5 M), shake at 37°C for 12h, magnetically separate, and wash to prepare the GSH fluorescent sensor;

[0023]Wherein, the GSH recognition probe is a nucleotide molecule containing a disulfide bond, and its base sequence is 5'-ACGAGTCA S-S TTT TGT TTG TTC CCC CCT TCT TTC TTA-3'; the sulfhydryl magnetic beads are Commodity purchased (Tianjin Beisi Le Chro...

Embodiment 2

[0025] A kind of method that adopts the GSH fluorescent sensor detection GSH based on controllable release technology that the present invention proposes is as follows:

[0026] (1) Add 10 μL of the GSH sample solution to be tested to the GSH fluorescent sensor based on the controllable release technology proposed by the present invention, dilute it to 100 μL with PBS buffer solution (PH=7.4), shake at a constant temperature of 37°C for 2 hours, and identify the probe and A specific molecular recognition reaction occurs between the target molecules GSH, which leads to the cutting of the disulfide bond, and the recognition probe containing the disulfide bond is detached from the surface of the gold nanocage, thereby opening the molecular gate that blocks the cage hole and dissipating the gold nanocage. The fluorescent molecule rhodamine B in the nanocage is released;

[0027] (2) Magnetic separation, collecting the supernatant and washing liquid for fluorescence detection, dete...

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Abstract

The invention provides a detection method for glutathione based on a controllable release technology. The method can be used for the fields, such as the early-stage diagnosis and treatment of a serious disease, food, pharmaceuticals, and bioscience. The provided method is used for detecting the glutathione on the basis of a fluorescence sensor of the controllable release technology. The method comprises the following steps: combining a gold nanoparticle cage with a glutathione recognition probe, forming a molecule gate with biological response on the surface of the gold nanoparticle cage through the static assembling function, using the specific molecular recognition reaction generated between the recognition probe and target molecules, causing the recognition probe to be separated from the surface of the gold nanoparticle cage, enabling the molecule gate to be opened, releasing fluorescence molecules in the gold nanoparticle cage, and realizing the strengthening and detection of a fluorescence signal. The method is capable of remarkably improving the detection sensitivity of the glutathione, and realizing the high-sensitive and high-selective detection for the glutathione.

Description

technical field [0001] The invention relates to a method for detecting glutathione (GSH), in particular to a method for detecting GSH which can be applied in many fields such as medical treatment, life science, food and medicine. Background technique [0002] Glutathione (GSH) is a tripeptide compound composed of glutamic acid, cysteine, and glycine. The sulfhydryl group in cysteine ​​residues makes it an important biologically active substance. GSH widely exists in animal and plant cells, and is the most abundant non-protein thiol in cells. The content of GSH in various tissues and cells of the human body and the relationship with various diseases have attracted much attention, and have been a hot spot of extensive research so far. GSH participates in a variety of biochemical reactions in organisms, and has very important physiological functions, such as the transport of amino acids, the synthesis of proteins and nucleic acids, and antioxidant effects. As an important ant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 王卫董颖杰王世颖申欣罗细亮
Owner 合肥龙之韵医药技术有限公司
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