Exosome for mass spectrometric flow cytometry detection and preparation method and application thereof

Abstract

The invention discloses an exosome for mass spectrometric flow cytometry detection and a preparation method and application thereof, and belongs to the field of nanobiological materials. The exosome comprises a heavy metal tag compound which can be used for mass spectrometric flow cytometry detection, wherein and the heavy metal tag compound is distributed in the lumen of the exosome. The preparation method comprises the following steps: 1) extracting the exosome: obtaining the exosome by gradient centrifugation, concentration and precipitation; or obtaining the exosome by ultra-high speed centrifugation; 2) introducing a heavy metal tag: introducing the heavy metal tag compound into the exosome by electroporation and incubation of the exosome obtained in the step 1) and the heavy metal tag compound; 3) obtaining the exosome with the heavy metal tag: removing the free heavy metal tag compound which is not encapsulated by the exosome by using a sedimentation kit or ultra-high speed centrifugation so as to obtain the exosome for mass spectrometric flow cytometry detection. The present application also provides the application of the exosome to realize high-throughput single cell tracing exosome uptake.

Description

technical field [0001] The invention belongs to the field of nano-biological materials, and in particular relates to an exosome used for mass spectrometer flow cytometry detection and its preparation method and application. Background technique [0002] Mass cytometry is currently the most advanced single-cell high-throughput proteomics technology in the world. It is the first to use metal elements as markers of flow-type antibodies and dyes, and use mass spectrometry to quantitatively detect labeled cells. The application of this new technology, on the one hand, greatly increased the number of flow detection channels to hundreds, increasing the amount of information obtained from a single sample; on the other hand, it avoided signal interference between channels, greatly simplified experimental design, and improved data reliability. The increase in the number of signaling molecules detected means more accurate observation and classification of phenotypes within cell popul...

AI Technical Summary

Problems solved by technology

Even though some studies have used traditional fluorescence-based cell flow cytometry to track the uptake of exosomes, due to the limitation of the number of fluorescent dye channels and the frequent occurrence of fluorescence leakage and cross-color, the traditional flow cytometry cannot be high Through-put resolution of cell types and high-throughput quantification of exosome uptake

Although the throughput of the mass spectrometry flow cytometer is very high, it can only detect 75-209 atomic weight elements, and naturally extracted exosomes do not contain these elements that can be taken up by the mass spectrometry flow cytometer, so the present invention Aim to construct an exosome with a heavy metal label compound, use mass spectrometry flow cytometry to track exosomes in vitro and in vivo, and establish a high-throughput method for tracking exosome uptake

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