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Fluorescence resonance energy transfer probe, preparation method and application thereof

A fluorescence resonance energy and probe technology, applied in biochemical equipment and methods, chemical instruments and methods, luminescent materials, etc., can solve the problems of loss of surgical treatment, high price, and high rate of missed diagnosis

Inactive Publication Date: 2016-09-21
XINXIANG MEDICAL UNIV
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AI Technical Summary

Problems solved by technology

Surgical resection is currently the most effective treatment, but most patients are diagnosed at an advanced stage, thus losing the opportunity for surgical treatment
At present, the main screening methods for liver cancer are alpha-fetoprotein AFP and real-time ultrasonography, but the missed diagnosis rate is high and the price is expensive

Method used

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  • Fluorescence resonance energy transfer probe, preparation method and application thereof
  • Fluorescence resonance energy transfer probe, preparation method and application thereof
  • Fluorescence resonance energy transfer probe, preparation method and application thereof

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Embodiment

[0025] 1 Materials and methods

[0026] 1.1 Cell lines and culture conditions

[0027] Human liver cancer cells 7402 and normal human liver cells LO-2 were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences, using RPMI-1640 medium containing 10% FBS and penicillin / streptomycin (penicillin 100 U / mL, streptomycin 100 μg / mL), At 37°C, 5% CO 2 cultured in a cell culture incubator.

[0028] 1.2 Main reagents and materials

[0029] 4-(N,N-diethylamino)salicylaldehyde, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl), N-hydroxysuccinyl Imine (NHS), N,N-dimethylformamide (DMF), petroleum ether (60-90°C), tetrahydrofuran (THF), chromatographically pure dimethyl sulfoxide (DMSO), etc. The rest of the chemical reagents used were of analytical grade. DMEM medium, fetal bovine serum (FBS), trypsin, penicillin, streptomycin, etc. are commercially available.

[0030] 1.3 Main Instruments

[0031] Ultraviolet-visible spectrophotometer (UV-2550...

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Abstract

The invention discloses a fluorescence resonance energy transfer probe, a preparation method and application thereof. According to the invention, the fluorescent group 7-(N, N-diethylamino)coumarin-3-carboxylic acid succinimidyl ester is connected to the quenching group 4-[4-(dimethylamino)benzeneazo]benzoic acid N-succinimidyl ester through a polypeptide chain able to be cut by the matrix metalloproteinase II (MMP-2) to construct the fluorescence resonance energy transfer probe. Human liver cancer 7402 cell and human normal liver cell LO-2 are adopted as the research object, and cell imaging experiments are carried out to detect the MMP-2 expression level of the 7402 cell and LO-2 cell. In vitro experiments (the fluorescence response of MMP-2, MMP-1 and other species to the probe) show the targeting of the probe to MMP-2, and the cell imaging result shows that the MMP-2 expression level of the human liver cancer 7402 cell is higher than the human normal liver cell LO-2.

Description

technical field [0001] The invention relates to a liver cancer cell detection technology, in particular to a fluorescence resonance energy transfer probe and a preparation method and application thereof. Background technique [0002] The incidence of liver cancer ranks fifth among malignant tumors in the world, and the death rate ranks third. It is one of the most common malignant tumors in my country. Liver cancer has an insidious onset, rapid development, and high mortality rate, which seriously threatens people's life and health. Surgical resection is currently the most effective treatment, but most patients are diagnosed at an advanced stage, thus losing the opportunity for surgical treatment. At present, the main screening methods for liver cancer are alpha-fetoprotein AFP and real-time ultrasonography, but the missed diagnosis rate is high and the price is expensive. Early diagnosis of liver cancer can greatly improve the 5-year survival rate of patients. Therefore,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C09K11/06C12Q1/37C12Q1/02
CPCC07K7/06C09K11/06C09K2211/1007C09K2211/1029C09K2211/1466C12Q1/37
Inventor 何广杰杨璐千新来原志庆李静冯思佳赵二趁
Owner XINXIANG MEDICAL UNIV
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