Simple and convenient exosome extraction method

Abstract

The invention discloses a simple and convenient exosome extraction method. The method comprises the following steps of (1) collecting exosome cell supernatant; (2) removing cell debris in the supernatant through centrifugation; (3) transferring the supernatant without the cell debris into an ultrafiltration tube, and enriching an exosome by using the ultrafiltration tube; (4) washing the exosome enriched in the ultrafiltration tube by using PBS; (5) salting out the washed exosome to obtain a relatively purified exosome; and (6) measuring the concentration of the purified exosome. According tothe method, the exosome is intercepted by using a filter membrane, and then high-molecular-weight protein is separated out through salting-out, so that the problem that the exosome is difficult to extract is solved; and the method is high in operability, and the problems that an exosome extraction reagent is expensive, operation is time-consuming, requirements for extraction equipment are high, and the product purity is low are truly solved. Therefore, by using the extraction method provided by the invention, the extraction method of the exosome is greatly simplified, and the extraction quality and extraction efficiency of the exosome are improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a simple method for extracting exosomes. Background technique [0002] Exosomes are tiny membrane vesicles that are secreted by most cells; in recent years, exosomes have shown more remarkable effects than cells in various disease models by virtue of their size and bimolecular membrane advantages. treatment effect. With the increasing research on exosomes, the research on exosomes derived from dendritic cells is also increasing. Existing extraction methods of exosomes mainly include the following types. [0003] 1. Ultracentrifugation method: The ultracentrifugation method is the most commonly used exosome extraction method. It uses low-speed centrifugation and high-speed centrifugation alternately to separate vesicle particles of similar size. The ultracentrifugation method is easy to operate and obtains a large number of vesicles, but the process is time-consuming an...

AI Technical Summary

Problems solved by technology

The ultracentrifugation method is easy to operate and obtains a large number of vesicles, but the process is time-consuming and the recovery rate is unstable; in addition, repeated centrifugation may cause damage to the vesicles, thereby reducing their quality

[0004] 2. PEG-base precipitation method: polyethylene glycol can be combined with hydrophobic proteins and lipid molecules to co-precipitate and extract exosomes, but the purity and recovery rate are low, there are many impurities (false positives), and the particle size is not uniform. Produce hard-to-remove polymers, mechanical force or chemical additives such as Tween-20 will destroy exosomes, etc.

The magnetic bead method has the advantages of high specificity, easy operation, and does not affect the integrity of exosomes, but the efficiency is low, and the biological activity of exosomes is easily affected by pH and salt concentration, which is not conducive to downstream experiments and is difficult to be widely used

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