A method for preparing an exosome

By combining tangential flow filtration, totipotent nuclease treatment, and gradient centrifugation, the exosome preparation process was optimized, solving the problem of difficulty in balancing exosome quality and yield in existing technologies, and achieving efficient and low-cost exosome preparation.

CN115710572BActive Publication Date: 2026-05-29苏州唯思尔康科技有限公司
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Patent Information

Application Number
CN202211552943.6
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2022-12-05
Publication Date
2026-05-29
Estimated Expiration
2042-12-05

AI Technical Summary

Technical Problem

Existing technologies struggle to simultaneously ensure both the quality and yield of exosomes, resulting in high production costs and low efficiency. Furthermore, the extracts often contain residual proteins and other structural vesicles, which impair the functional performance of exosomes.

Method used

The cell supernatant was concentrated using tangential flow filtration, combined with totipotent nuclease treatment, density gradient centrifugation, and ultracentrifugation. By optimizing the centrifugal force and buffer composition, exosomes were separated and purified, ensuring membrane structure integrity and high yield.

Benefits of technology

High-quality, high-purity exosomes with intact membrane structures, high natural activity, improved yield, and reduced production costs were obtained, making them suitable for industrial preparation.

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Abstract

The present application relates to a preparation method of exosomes, comprising the following steps: 1) concentrating cell supernatant to obtain a concentrated solution by using a tangential flow filtration method; 2) removing nucleic acids from the concentrated solution by using a universal nuclease and resuspending to obtain a crude extract; 3) performing density gradient centrifugation on the crude extract, and collecting the exosome layer; 4) performing low-speed centrifugation on the exosome layer under a centrifugal force less than or equal to 50000g, and removing the precipitate to obtain a preliminary purified solution; and 5) performing ultracentrifugation on the preliminary purified solution under a centrifugal force greater than or equal to 100000g, and the obtained precipitate is the exosomes. The exosomes obtained by the method have a complete and un-deformed membrane structure, higher purity and particle concentration, and better quality compared with exosomes prepared by existing methods. Meanwhile, the method has high yield and low cost, and helps to realize the industrialized preparation of exosomes, and provides strong support for targeted drug delivery research using exosomes as carriers.
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