title: PEG precipitation coupled with chromatography is a new and sufficient method for the purification of botulinum neurotoxin type B [corrected].
abstract: Clostridium botulinum neurotoxins are used to treat a variety of neuro-muscular disorders, as well as in cosmetology. The increased demand requires efficient methods for the production and purification of these toxins. In this study, a new purification process was developed for purifying type B neurotoxin. The kinetics of C.botulinum strain growth and neurotoxin production were determined for maximum yield of toxin. The neurotoxin was purified by polyethylene glycol (PEG) precipitation and chromatography. Based on design of full factorial experiment, 20% (w/v) PEG-6000, 4 °C, pH 5.0 and 0.3 M NaCl were optimal conditions to obtain a high recovery rate of 87% for the type B neurotoxin complex, as indicated by a purification factor of 61.5 fold. Furthermore, residual bacterial cells, impurity proteins and some nucleic acids were removed by PEG precipitation. The following purification of neurotoxin was accomplished by two chromatography techniques using Sephacryl™ S-100 and phenyl HP columns. The neurotoxin was recovered with an overall yield of 21.5% and the purification factor increased to 216.7 fold. In addition, a mouse bioassay determined the purified neurotoxin complex possessed a specific toxicity (LD(50)) of 4.095 ng/kg.
authors: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Fengtai District, Beijing, People's Republic of China.
authors: Yao
authors: Y
authors: Zhao
authors: Lin
authors: L
authors: Kang
authors: Shan
authors: S
authors: Gao
authors: Xing
authors: X
authors: Gao
authors: Wenwen
authors: W
authors: Xin
authors: Jinglin
authors: J
authors: Wang
mesh_terms: D000818:Animals; D001905:Botulinum Toxins / Q000302:isolation & purification* / Q000633:toxicity; D019274:Botulinum Toxins, Type A; D002853:Chromatography, Liquid / Q000379:methods*; D051379:Mice; D011092:Polyethylene Glycols / Q000737:chemistry*
chemical_list: C096323:rimabotulinumtoxinB; D011092:Polyethylene Glycols; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A