Extractant used for extracting extracellular vesicle in body fluid and extracting method thereof

Abstract

The invention relates to an extractant used for extracting extracellular vesicle in body fluid and an extracting method thereof. The extractant is prepared from inorganic salt and a water-soluble polymer, the inorganic salt is selected from one or more of ammonium sulfate, calcium chloride, sodium chloride, magnesium chloride or sodium citrate, and the water-soluble polymer is selected from one ofpolyethylene glycol PEG, methylcellulose MC and polyethyleneimine PEI. The extracting method comprises the steps that the extractant is mixed with the body fluid to form a to-be-extracted system, andafter salting-out and centrifugation, the extracellular vesicle can be extracted out. The adopted salting-out method for extracting the extracellular vesicle has the three advantages that 1), the yield is high and is about 70%; 2) the operation is quick and convenient, the extracellular vesicle can be extracted out within 30-45 minutes; 3) the method is not limited by the size of a sample and issuitable for extracting of samples with large-sizes.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an extractant for extracting extracellular vesicles in body fluids and an extraction method thereof. Background technique [0002] Living cells will continuously secrete extracellular vesicles, the main types of which include exosomes (50-100nm in size) and microvesicles (20-1000nm in size). In humans, endogenous extracellular vesicles have been shown to play roles in coagulation, intercellular signaling, and waste management. At the same time, extracellular vesicles have great potential in the diagnosis and treatment of diseases because they contain nucleic acids, proteins, lipids and other biomolecules. [0003] Currently, methods for isolating extracellular vesicles from body fluids include ultracentrifugation, immunocapture, ultrafiltration, polymer precipitation, and electrostatic adsorption. Among them, the equipment required for ultracentrifugation is expensive, the operatio...

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Problems solved by technology

Among them, the equipment required for ultracentrifugation is expensive, the operation is complicated, and the recovery rate is not high.

The immunocapture method uses specific antibodies to identify and separate extracellular vesicles, which has high specificity, but it is expensive and not suitable for processing large volume samples, and it is easy to lose vesicles with low antigen expression

Ultrafiltration can quickly separate vesicles in body fluids based on particle size or molecular weight, but vesicles may be revived by ultrafiltration membranes with micron or nanopore sizes and affect the final extraction efficiency

The polymer precipitation method has a high yield, but there are a lot of protein impurities in the product, and the precipitation process needs to be incubated for a period of time (0.5 hours to overnight), and the time required to complete an operation is still very long

The electrostatic adsorption method does not require large-scale equipment, the device is simple, and the product is of high purity, but it is not suitable for processing large-volume samples

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