Hennessey J P, Oswald C B, Dong Z, Lewis J A, Sitrin R D
Bioprocess and Bioanalytical Research, Merck Research Laboratories, West Point, PA 19486, USA.
Vaccine. 1999 Jul 16;17(22):2830-5. doi: 10.1016/s0264-410x(99)00095-x.
Manufacture of VAQTA, an inactivated hepatitis A virus vaccine, includes extensive purification of the intact virus particle to remove endogenous components from the host cell culture lysate as well as compounds introduced in the upstream purification process. Analysis of the final purified hepatitis A virus product by SDS-PAGE prior to inactivation shows that greater than 95% of the protein in the preparation is found in four protein bands, which have been confirmed to be hepatitis A virus capsid proteins VP0, VP1, VP2 and VP3 based on Western blot and mass spectrometry analyses. Validation of the manufacturing process and direct analysis of the final product were used to demonstrate that no other specific host cell-derived components are detected and that process residuals are all below the limits of detection of the assays used. Establishment of a rigorous standard of high purity for this product was pursued to minimize the impact of impurities during clinical development of this product and will facilitate the incorporation of this product into combination vaccines.
甲肝灭活疫苗VAQTA的生产包括对完整病毒颗粒进行广泛纯化，以去除宿主细胞培养裂解物中的内源性成分以及上游纯化过程中引入的化合物。在灭活之前通过SDS-PAGE对最终纯化的甲型肝炎病毒产品进行分析表明，制剂中超过95%的蛋白质存在于四条蛋白带中，基于蛋白质印迹和质谱分析，已证实这四条蛋白带为甲型肝炎病毒衣壳蛋白VP0、VP1、VP2和VP3。生产工艺的验证和最终产品的直接分析用于证明未检测到其他特定的宿主细胞衍生成分，并且工艺残留均低于所用检测方法的检测限。为该产品建立严格的高纯度标准，以尽量减少杂质在该产品临床开发过程中的影响，并将有助于将该产品纳入联合疫苗。
