Hagen A J, Aboud R A, DePhillips P A, Oliver C N, Orella C J, Sitrin R D
Vaccine BioProcess R&D Department, Merck Research Laboratories, West Point, PA 19486, USA.
Biotechnol Appl Biochem. 1996 Jun;23(3):209-15.
The development of the purification process for VAQTA, which results in a highly purified inactivated hepatitis A vaccine, was driven by modifications in the cell-culture and harvest methods which permit hepatitis A virus propagation to support large-scale manufacture. The starting material for the purification was initially a concentrated cell pellet scraped from roller bottles. However, when the cell-culture method was scaled up to use high-surface-area Nunc cell factories or Costar cubes, the early steps in the process had to be modified to handle large volumes of dilute lysate. Membrane concentration was used at first, and a highly purified vaccine was prepared, but virus-poly(nucleic acid) complexes were formed, which reduced the yields in later processing steps. The introduction of a nuclease digestion immediately after harvest followed by capture chromatography on an anion-exchange column eliminated the formation of these complexes and resulted in more consistent performance and higher yields of downstream operations.
VAQTA纯化工艺的发展，产生了一种高度纯化的甲型肝炎灭活疫苗，这一发展得益于细胞培养和收获方法的改进，这些改进使得甲型肝炎病毒能够繁殖以支持大规模生产。纯化的起始原料最初是从滚瓶上刮下的浓缩细胞沉淀。然而，当细胞培养方法扩大规模以使用高表面积的Nunc细胞工厂或Costar培养盒时，该工艺的早期步骤必须进行修改以处理大量稀释裂解物。最初使用膜浓缩法制备了高度纯化的疫苗，但形成了病毒 - 多（核酸）复合物，这降低了后续加工步骤的产量。收获后立即引入核酸酶消化，然后在阴离子交换柱上进行捕获色谱，消除了这些复合物的形成，并导致下游操作具有更一致的性能和更高的产量。
