Procedures for large-scale production and purification of Clostridium botulinum C1 toxin for preparation of toxoid
Procedures for large-scale production and purification of Clostridium botulinum C1 toxin for preparation of toxoid
复制标题
用于制备类毒素的肉毒梭菌C1毒素的大规模生产和纯化程序
DOI：
10.1111/j.1574-6968.1985.tb00983.x
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发表时间：
1985
影响因子：
2.1
通讯作者：
G. Sakaguchi
中科院分区：
文献类型：
--
作者：
H. Kurazono;K. Shimozawa;M. Hosokawa;G. Sakaguchi
含碳酸钙的强化熟肉培养基（CaCO 3-FCM）支持C型肉毒梭菌菌株的毒素产生水平达到2 × 106小鼠i.p.LD50/ml。C1毒素通过以下顺序步骤纯化：在RNA存在下从5倍稀释的培养物上清液中酸沉淀，通过透析进行第二次酸沉淀，通过鱼精蛋白处理去除RNA，通过Amicon PM-30膜超滤去除过量的鱼精蛋白和缓冲液，磺丙基-Sephadex层析和Sephadex G-200凝胶过滤。通过这些方法，从一批600-ml培养物中恒定地获得25 mg或更多的高度纯化的C1毒素。
Fortified cooked meat medium containing calcium carbonate (CaCO3-FCM) supported toxin production of a strain of Clostridium botulinum type C to a level of 2 × 106 mouse i.p. LD50/ml. C1 toxin was purified by sequential steps of acid precipitation from 5-fold diluted culture supernatant in the presence of RNA, 2nd acid precipitation by dialysis, removal of RNA by protamine treatment, removal of excess protamine and bufferisation by ultrafiltration through Amicon PM-30 membrane, sulphopropyl-Sephadex chromatography, and Sephadex G-200 gel filtration. By these procedures, 25 mg or more of highly purified C1 toxin was constantly obtained from a lot of 600-ml culture.