Extracellular vesicle enrichment detection method

Abstract

The invention belongs to the technical field of medical detection, and discloses an extracellular vesicle enrichment detection method, which is characterized in that exosome enrichment detection is performed after DNA tetrahedron synthesis, magnetic bead combination and exosome extraction. According to the method, exosome surface specific protein is utilized, on the basis of a DNA origami aptamer and immunoaffinity magnetic bead space recognition dual recognition and capture method, a DNA fluorescent probe is further combined, so that rapid separation and capture of the exosome are realized, and the method can be used for quantitatively detecting the exosome. The invention is suitable for being used as a kit to realize enrichment, separation and detection of exosomes, and has very important significance in wide application in the fields of disease diagnosis, pathological research, new drug development and the like.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and relates to an exosome enrichment detection method, in particular to an extracellular vesicle enrichment detection method. Background technique [0002] Exosomes refer to small vesicles secreted by cells, that is, extracellular vesicles, with a diameter of mostly between 30-150nm. Exosomes contain proteins, lipids, nucleic acids and other substances, which can be received by recipient cells to realize material transportation and information transmission between cells. Almost all biological cells can release exosomes in various states, and even some microorganisms can release exosomes for signal transmission. Exosomes contain many key proteins and special genetic materials, which play an important role in antigen presentation, immune regulation, tissue development, intercellular communication, and mediate tumor development and development in cell communication and epigenetic regulati...

AI Technical Summary

Problems solved by technology

However, during the ultrafiltration process of ultrafiltration, exosomes may block the filter pores, resulting in shortened membrane life and low separation efficiency. In addition, exosome membranes will also adhere to each other, resulting in low separation yield and even False detection results occur; polyethylene glycol in the PEG-base precipitation method can compete with free water molecules, so that less soluble molecules or exosomes are precipitated from the solution. There are also some problems in this method: such as exosomes The purity and recovery rate of exosomes are low, and mechanical force or chemical additives can damage exosomes, etc.; while the magnetic bead immunoassay method can obtain high-purity exosomes due to the high specificity of immunoaffinity technology without affecting The advantages of complete exosome morphology are the preferred method for enriching and characterizing unique exosomes, but this method is inefficient, and the biological activity of exosome contents is easily affected by pH and salt concentration, which is not conducive to the downstream experiments

[0005] DNA origami technology is to repeatedly fold long-chain DNA to construct various tiny 3D structures, including micro-biosensors and drug delivery containers. At present, this technology has been applied to exosome detection, but the structure and construction rules are complicated. It is difficult to design and visualize the double-helix chain of DNA molecules bent into a specific shape. The existing technology focuses more on the labeling and detection of exosomes, and its origami structure is not effective in the enrichment and separation of exosomes.

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