Fluorescent enzyme-linked immunoassay method for detection of galectin-4

An immunoassay, luciferase technology, applied in the direction of material analysis, analysis of materials, fluorescence/phosphorescence, etc. by optical means, can solve the problems of complex assembly process, high cost of carrier, limited application, etc., to achieve sensitive response and improve catalytic activity. , high sensitivity and stability

Active Publication Date: 2016-12-21
FUZHOU HOSPITAL FOR INFECTIOUS DISEASE
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Problems solved by technology

However, some of these immobilization methods have shortcomings such as long time, covalent modification or c

Method used

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  • Fluorescent enzyme-linked immunoassay method for detection of galectin-4
  • Fluorescent enzyme-linked immunoassay method for detection of galectin-4
  • Fluorescent enzyme-linked immunoassay method for detection of galectin-4

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Embodiment 1

[0034] 1. Synthesis of gold nanoclusters

[0035] Before the synthesis, soak the glassware used in aqua regia, then rinse with a large amount of water, and finally rinse with ultrapure water for use. Accurately pipette 5 ml of 4mM chloroauric acid solution and 5 ml of 8mM glutathione solution in a round bottom flask at room temperature and mix well. The flask was then transferred to a 70°C oil bath for 24 hours of reaction. After cooling to room temperature, transfer the reaction solution to a 1000Da molecular weight cut-off dialysis bag and dialyze with ultrapure water for 24 hours to remove unreacted chloroauric acid and glutathione molecules, and centrifuge the obtained dialysate at 10,000rpm for 10 minutes to remove the precipitate , to obtain a gold nanocluster solution, which was stored in a refrigerator at 4°C and protected from light until use. The spectra of the synthesized gold nanoclusters are shown in figure 2 As shown, when excited at 375nm, gold nanoclusters ...

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Abstract

The invention relates to a fluorescent enzyme-linked immunoassay method for detection of galectin-4. According to the method, glucose oxidase on an enzyme labeled complex combined through a target substance catalyzes glucose to generate hydrogen peroxide, then a ferrous ion solution is added to generate reactive oxygen for quenching fluorescence of a gold nanocluster solution, and fluorescence change is determined by a fluorescence spectrometer for quantitative detection of a target molecule. Fluorescent gold nanoclusters used in the method have stable luminescence and are sensitive to response to the reactive oxygen, and are more stable in controllability and more convenient to operate when used as a detection probe. The detection system constructed by the method has the advantages of simple synthesis, low cost, and high sensitivity and stability, and is expected to be more widely applied and extended in clinical biomolecular detection.

Description

(1) Technical field [0001] The invention relates to a fluorescent enzyme-linked immunoassay method for detecting galectin-4. (2) Background technology [0002] Galectin-4 (galectin-4) is a β-galactoside-binding protein, which is a member of the lectin family. It has a CRD domain and has a high affinity with glycoproteins and glycolipids containing β-galactoside. , plays a specific adhesion role between cells and cells, and between cells and matrix, and is closely related to tumor metastasis, invasion, growth and adhesion. Studies have shown that the abnormal expression of galectin-4 is related to some cancers, for example, the content of galectin-4 in the serum of patients with metastatic colon cancer and breast cancer is significantly increased; in addition, in pancreatic cancer, hepatocellular carcinoma, gastric cancer The content of galectin-4 was also up-regulated in patients. Because galectin-4 plays an important role in cancer cell metastasis and growth, it is expect...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/532G01N21/64
CPCG01N21/6428G01N33/532G01N33/68G01N2021/6439
Inventor 刘景丰曾永毅刘小龙张晓龙郑爱仙廖乃顺
Owner FUZHOU HOSPITAL FOR INFECTIOUS DISEASE
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