Method for preparing nano fluorescent probe with high bio-safety

Abstract

The invention discloses a method for preparing a nano fluorescent probe with high bio-safety. Fluorescent protein is wrapped by using a natural macromolecular chitosan material with extremely high bio-safety to improve the stability of the fluorescent protein and expand the using field and range of the fluorescent protein. The preparation method comprises the steps of adsorption and cross-linking. The method is quite simple and feasible and extremely high in wrapping efficiency; and the obtained nano particles have high stability, dispersibility and bio-safety, can protect the protein from being hydrolyzed by enzymes and denatured, and also have excellent performance of protecting the fluorescence from being quenched and blanched. In-vitro experiments show that the material can successfully enter cells and is applied to molecule level as the fluorescent probe with high bio-safety.

Description

technical field [0001] The invention relates to a preparation method of a nano fluorescent probe with good biological safety, belonging to the fields of polymer nanomaterials and biochemistry. Background technique [0002] Fluorescent probes have been widely used in biomedical fields, including imaging, sensing, and diagnosis. However, the biological safety of traditional fluorescent probes has been controversial. For example, fluorescent probes prepared with traditional organic dyes are still used today. However, due to serious problems in stability and safety of organic dyes, their applications in the biomedical field have been gradually replaced by new nanoprobes, such as quantum dots. However, most quantum dot materials (such as CdSe, etc.) contain heavy metal elements , the toxicity of its heavy metals is a potential hidden danger, which limits its application. Although the risk of releasing heavy metal ions can be reduced by encapsulation by silica gel or polymer, onc...

AI Technical Summary

Problems solved by technology

[0002] Fluorescent probes have been widely used in biomedical fields, including imaging, sensing, and diagnosis. However, the biological safety of traditional fluorescent probes has been controversial. For example, fluorescent probes prepared with traditional organic dyes are still used today. However, due to serious problems in stability and safety of organic dyes, their applications in the biomedical field have been gradually replaced by new nanoprobes, such as quantum dots, but most quantum dot materials (such as CdSe, etc.) contain heavy metal elements , the toxicity of its heavy metals is a potential hidden danger, which limits its application. Although the risk of releasing heavy metal ions can be reduced by encapsulating it with silica gel or polymer, once it leaks, there will be a huge safety hazard, which limits its use as a fluorescent probe in biological In vivo application

[0006] The current technology only uses traditional methods such as emulsification cross-linking or complex coacervation to prepare chitosan nanoparticles, and wraps proteins and other drugs through physical embedding or adsorption. This method has a very low wrapping efficiency (generally less than 60 %), and the prepared nanoparticles are large, some even reach the micron level, so the dispersion and stability of the nanoparticles are not good, and they cannot be stored for a long time, otherwise serious agglomeration will occur

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