METHOD FOR PREPARING CLOSTRIDIUM BOTULINUM NEUROTOXIN WITH BIOACTIVITY AND MULTILAYERED MULTIPHASE NANOPARTICLES

摘要

The present invention relates to the field of protein production and transdermal delivery, and discloses a method for producing Clostridium botulinum neurotoxin having bioactivity and multi-layered multiphase nanoparticles. The method comprises fermenting Clostridium botulinum bacteria in a fermentation medium free of animal-derived components, bringing the fermentation medium into contact with the anion exchange medium slurry, and obtaining a supernatant containing Clostridium botulinum neurotoxin through centrifugation do. In this method, the supernatant is dialyzed to collect a dialysate containing Clostridium botulinum neurotoxin, the dialysate is contacted with an anion exchange chromatography column, and the eluate collected from the anion exchange chromatography column is contacted with a cation exchange chromatography column to obtain the eluate. It further comprises the step of collecting. The nanoparticles include an aqueous phase core or oil phase core as an innermost layer, an aqueous phase layer, an oil phase layer and a cream layer as the outermost layer. The aqueous phase and the oil phase are alternately encapsulated to form multi-layered multiphase nanoparticles. Biomolecules include active molecules such as Clostridium botulinum neurotoxin and/or hyaluronic acid.