Tan L U, Fahim R E, Jackson G, Phillips K, Wah P, Alkema D, Zobrist G, Herbert A, Boux L, Chong P
Connaught Laboratories Limited, Willodale, Ontario, Canada.
Mol Immunol. 1991 Mar;28(3):251-5. doi: 10.1016/0161-5890(91)90070-z.
A novel process for preparing non-pyrogenic toxoids of pertussis toxin (PT) and filamentous hemagglutinin (FHA) is described. The process consists of chromatographies on perlite then on hydroxylapatite. Purification yields for PT and FHA are 62 and 68%, respectively. The purification process takes advantage of the novel use of perlite (a filter aid) for the simultaneous purification of PT and FHA. The hydroxylapatite, in addition to removing the remaining contaminants, also concentrates the antigens. The resulting PT and FHA are approximately 95% pure, and are non-pyrogenic as judged by the rabbit pyrogen test. The purification process is simple, inexpensive, and does not use blood components or toxic substances. The mild conditions in which the PT and FHA are purified ensure the recovery of native protein. The purified PT and FHA are detoxified in the presence of glycerol using glutaraldehyde and formaldehyde, respectively, to produce antigenic components of an acellular pertussis vaccine. The final PT and FHA toxoids are immunogenic in guinea-pigs and have been shown to be protective in the mouse intracerebral challenge test.
描述了一种制备百日咳毒素（PT）和丝状血凝素（FHA）非热原性类毒素的新方法。该方法包括先在珍珠岩上进行色谱分离，然后在羟基磷灰石上进行色谱分离。PT和FHA的纯化产率分别为62%和68%。该纯化过程利用了珍珠岩（一种助滤剂）的新用途，可同时纯化PT和FHA。羟基磷灰石除了去除残留污染物外，还能浓缩抗原。所得的PT和FHA纯度约为95%，通过兔热原试验判断为非热原性。该纯化过程简单、廉价，且不使用血液成分或有毒物质。纯化PT和FHA的温和条件确保了天然蛋白质的回收。纯化后的PT和FHA分别在甘油存在下使用戊二醛和甲醛进行解毒，以产生无细胞百日咳疫苗的抗原成分。最终的PT和FHA类毒素在豚鼠中具有免疫原性，并已在小鼠脑内攻毒试验中显示出保护作用。
