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    • 38. 发明授权
    • Multilayer nano-cell containing biomolecules
    • US10851363B1
    • 2020-12-01
    • US16419508
    • 2019-05-22
    • Boke Zhang
    • Boke Zhang
    • C12N9/52A61K9/51A61K31/728B01D15/36B01D61/24A61K38/00
    • Method for preparing botulinum neurotoxin and nanoparticle thereof are provided. The method includes fermenting bacteria Clostridium botulinum in a fermentation media free of animal-derived ingredients and contacting the fermentation media with an anion exchange media slurry and obtaining a supernatant including the botulinum neurotoxin by centrifugation. The method further includes dialyzing the supernatant and collecting a dialyzed solution including the botulinum neurotoxin, contacting the dialyzed solution with an anion exchange chromatography column, contacting an elute collected from the anion exchange chromatography column with a cation exchange chromatography column, and collecting an elute. The nanoparticle includes multi-layer including an innermost water phase core including biomolecules encapsulated by an oil phase layer, thereby forming a water-in-oil structure, water phase layers; oil phase layers; and an outmost cream layer. The water phase layers and the oil phase layers alternatively encapsulate the water-in-oil structure. The biomolecules include botulinum neurotoxin and/or hyaluronic acid.