A kind of extracellular vesicle purification material and purification method
A purification method and vesicle technology, applied in the biological field, can solve the problems of low separation method, low efficiency, impurity of purified extracellular vesicle products, etc., and achieve the effect of flexible carrier
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[0093] Specifically, the preparation method of the nano-zirconia microspheres comprises the following steps:
[0094] (1) Preparation of zirconium dioxide precursor: mix zirconium dioxide powder, the first part of concentrated sulfuric acid and ammonium sulfate and heat to dissolve, then add the second part of concentrated sulfuric acid after cooling to room temperature, and mix well to form zirconium dioxide Precursor; preferably, the weight volume ratio of the zirconia powder, the first part of concentrated sulfuric acid, ammonium sulfate and the second part of concentrated sulfuric acid is 0.09-0.11g: 9-11ml: 2-4g: 18-22ml;
[0095] (2) Preparation of liquid A: Povidone and sodium dodecylbenzene sulfonate are dissolved in deionized water and fully stirred to dissolve to form liquid A; preferably, povidone, sodium dodecylbenzene sulfonate and deionized water The weight-to-volume ratio of ionized water is 0.05g: 2g: 100ml;
[0096] (3) Preparation of nano-halogen dioxide mic...
Embodiment 1
[0137] This example provides a method for purifying extracellular vesicles: using nano-zirconia microspheres to purify extracellular vesicles;
[0138] The preparation method of nano zirconia microspheres is as follows:
[0139] (1) Self-made zirconia precursor: Accurately weigh 0.10 g of zirconia powder, put it into a 150 mL beaker, add 10 mL of concentrated sulfuric acid and 3 g of ammonium sulfate, cover with a watch glass, heat and dissolve on an electric furnace; After the sample is completely dissolved, take it out and cool it slightly, put it into a 200 mL volumetric flask, and add 20 mL of concentrated sulfuric acid;
[0140] (2) Liquid A: 0.05g of povidone and 2g of sodium dodecylbenzenesulfonate are dissolved in 100ml of deionized water, fully stirred and dissolved for later use;
[0141] (3) Preparation of nano-Zirconium dioxide microspheres: Add 70ml of liquid A into a stirring bottle, blow in nitrogen, stir at 30°C and 800rpm for 15 minutes, slowly add 30ml of zi...
Embodiment 2
[0143] This example provides a purification method of extracellular vesicles: put the sample into the buffer, and then put nano-titanium dioxide microspheres that can reversibly bind to phosphatidylserine, and the extracellular vesicles are adsorbed on the surface of the microspheres , while other impurities are not adsorbed and removed, then the microspheres adsorbed with extracellular vesicles are washed with washing solution, and finally washed with EB eluent to obtain purified extracellular vesicles;
[0144] The preparation method of nano titanium dioxide microsphere is as follows:
[0145] (1) Self-made titanium dioxide precursor: Dissolve 8ml tetrabutyl titanate in 4ml absolute ethanol, slowly add 100ml deionized water and stir thoroughly, then add 1.4ml concentrated hydrochloric acid, seal and place at room temperature for 48 hours to obtain a light yellow transparent liquid;
[0146] (2) Liquid A: 0.05g of povidone and 2g of sodium dodecylbenzenesulfonate are dissolve...
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