Latham W C, Michelsen C B, Edsall G
Appl Microbiol. 1967 May;15(3):616-21. doi: 10.1128/am.15.3.616-621.1967.
Sephadex gel filtration can be employed as a preparative procedure for the purification of both tetanus and diphtheria toxoids. A toxoid purification sequence is described in the text. By utilizing the described methods and columns, up to 100,000 human doses of diphtheria toxoid could be processed in a single operation. The method has given an 80% yield of diphtheria toxoid with a purity of 1,900 Lf per mg of N. The analysis of the material by immunodiffusion tests showed that a marked increase in purity was achieved. Antigenicity tests demonstrated that there was no significant difference in antigenic potency between the parent toxoid and its purified fraction. Factors limiting the effective separation of tetanus toxoid by gel filtration are discussed. The construction of the columns used is described in detail, as well as packing procedures and column characteristics such as bed volume, void volume, sample size, and flow rate.
葡聚糖凝胶过滤可作为制备破伤风类毒素和白喉类毒素的纯化方法。文中描述了类毒素的纯化步骤。通过使用所述方法和柱，单次操作可处理多达100,000人份剂量的白喉类毒素。该方法白喉类毒素的产率为80%，纯度为每毫克氮1900Lf。通过免疫扩散试验对材料进行分析表明，纯度有显著提高。抗原性试验表明，母类毒素与其纯化部分之间的抗原效力没有显著差异。讨论了凝胶过滤法有效分离破伤风类毒素的限制因素。详细描述了所用柱的构造，以及填充程序和柱的特性，如床体积、空隙体积、样品量和流速。
