Exosome separation and purification device and separation and purification method thereof

Abstract

The invention relates to an exosome separation and purification device and a separation and purification method thereof. The device comprises an upper micro-fluidic chip, a filter membrane and a lowermicro-fluidic chip, and the filter membrane is arranged between the upper micro-fluidic chip and the lower micro-fluidic chip. Sample liquid is injected into the upper micro-fluidic chip through oneinlet of the upper micro-fluidic chip, and after the sample liquid is filtered through the filtering membrane, an exosome is obtained from one inlet of the lower micro-fluidic chip. According to the method disclosed by the invention, impurity protein in the exosome can be removed, the whole exosome detection and analysis time of the technology is less than 1 hour, and the technology is stable. A sample obtained by the device can be directly subjected to mass spectrometry, and the main signals of a graph obtained are consistent with those of a mass spectrum detection graph obtained after exosome cyst separation and purification through an existing standard ultracentrifugation method. The method has the advantages of being low in cost, small in amount of samples needed, rapid, convenient touse, accurate, stable, capable of avoiding the defects that time and resources are consumed in the ultracentrifugation technology and the like and wide in biological and medical application prospect.

Description

technical field [0001] The invention relates to an exosome separation and purification device and a separation and purification method thereof, belonging to medical experiment analysis methods. Background technique [0002] Exosomes are a kind of extracellular vesicles (Extracellular Vesicles, EV, the size of extracellular vesicles is about 30-1000nm), and its diameter is about 30-150nm. Rich in membrane proteins. Exosome vesicles contain proteins, nucleic acids (miRNA, mRNA, etc.), membrane proteins, or lipids. [0003] Microfluidics integrates the basic operation units such as sample preparation, reaction, separation, and detection in the biological, chemical, and medical analysis processes into a micron-scale chip to automatically complete the entire analysis process. Due to its great potential in the fields of biology, chemistry, and medicine, it has developed into a new research field interdisciplinary in biology, chemistry, medicine, fluid, electronics, materials, an...

AI Technical Summary

Problems solved by technology

[0004] Ultracentrifugation is currently the "gold standard" of exosome isolation methods. However, ultracentrifugation technology has the disadvantages of time-consuming, expensive, and large sample loss. Membrane filtration technology can separate and purify exosomes. When the sample size Larger use of filtration membranes to separate and purify exosome vesicles is easy to block or block, and whether exosomes pass through the filtration membrane is easy to adhere and cannot be eluted

[0005] In the prior art, the Chinese invention patent No. ZL2016105973232, titled "Extraction Method of Exosomes in Human Body Fluids" discloses "a. Preparation of body fluid samples: first extract human body fluids under sterile conditions and dilute them with PBS buffer. Then preliminarily remove the cell components and cell fragments in the body fluid by centrifugal screening, and make a body fluid sample for later use; b, body fluid sample purification: filter the body fluid sample in step a through a filter membrane, and further remove cell debris and other impurities in the body fluid , let it stand for 10-15 minutes, and keep the precipitate for later use; c, exosome extraction: suspend the precipitate obtained in step b with PBS buffer, suspend the exosome in the upper layer of the liquid, and then use a sterile needle Absorb the liquid containing exosomes in the upper layer and store it at -80°C for future use.” Although this patent discloses that the body fluid sample is filtered and extracted through a filter membrane, the accuracy of the mass spectrometry detection results after filtration cannot meet the requirements. Filtration and treatment methods are also not up to the requirements, and cannot meet the requirements of replacing ultracentrifugation for experimental detection

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