Fluorescent probe for detecting biologic thiol and preparation method and usage method thereof

Abstract

The invention discloses a fluorescent probe for detecting a biologic thiol and a preparation method and a usage method thereof. The fluorescent probe for detecting the biologic thiol consists of two parts: namely, a 2, 4-bi-nitrobenzene sulfonyl group which is a recognizing group and a boron difluoride-dipyrryl methane (BODIPY) derivative which is an information reporting functional group. The molecule of the probe can simply and quickly enter a living cell, generates a specificity reaction with the thiol in the cell, and causes to obviously enhance fluorescence intensity, so that the fluorescent probe further can be used for the fluorescence detection and the imaging of the active thiol in the living cell. The fluorescent probe has good stability, can be stored and used for a long time, is applicable to various environments in which the living cell grows, and has higher detection sensitivity for the thiol, strong anti-interference capability, excellent selectivity and no action on other common biologic interfering molecules. The fluorescent probe can simply enter the living cell and a living tissue; the single recognition of the thiol in a biologic system can be achieved effectively; and therefore, the fluorescent probe can be used for the fluorescence imaging of the living cell.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a fluorescent probe for detecting biological sulfhydryl groups, a preparation method and a use method thereof. Background technique [0002] Sulfhydryl is the group with the highest chemical activity in cells, an important component of many proteins and small molecules in organisms, and plays an important role in the antioxidant system of cells. The content of sulfhydryl molecules is directly related to various diseases, such as cancer, Parkinson's disease, cardiovascular disease, etc. Therefore, it is of great significance to develop a rapid, sensitive and simple method for detecting thiols in biochemistry and clinical chemistry. Traditional detection methods include mass spectrometry, high performance liquid chromatography, electrochemical methods, and fluorescence methods. Due to its high sensitivity and wide dynamic response range, more importantly, fluore...

AI Technical Summary

Problems solved by technology

For example, the developed aryl halides, dansylaziridines, pyrenes, etc. have good selectivity and sensitivity, and the detection limit for sulfhydryl groups is generally around nanomolar (nmol), but these probes themselves have Strong fluorescence, which will cause low signal-to-noise ratio when used in live cell imaging; benzofuransulfonyl halide probes, although their autofluorescence is weak, need to react with sulfhydryl groups under alkaline and high temperature conditions, and cannot Applied to fluorescence imaging of living cells; acetyl halide derivatives, such as sulfhydryl derivative reagents, can react rapidly with sulfhydryl groups at room temperature and physiological pH, but their photostability is poor, and they are easy to lose fluorophores, so it is difficult to meet the requirements of biological detection and fluorescence. Imaging Requirements

So far, highly sensitive and highly responsive probes that can be truly used for intracellular thiol localization imaging in living cells are still relatively rare

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