Onions David, Egan William, Jarrett Ruth, Novicki Deborah, Gregersen Jens-Peter
BioReliance Corporation, Rockville, USA.
Biologicals. 2010 Sep;38(5):544-51. doi: 10.1016/j.biologicals.2010.04.003.
Cell culture-based production methods may assist in meeting increasing demand for seasonal influenza vaccines and developing production flexibility required for addressing influenza pandemics. MDCK-33016PF cells are used in propagation of a cell-based seasonal influenza vaccine (Optaflu); but, like most continuous cell lines, can grow in immunocompromised mice to produce tumors. It is, therefore, essential that no residual cells remain within the vaccine, that cell lysates or DNA are not oncogenic, and that the cell substrate does not contain oncogenic viruses or oncogenic DNA. Multiple, redundant processes ensure the safety of influenza vaccines produced in MDCK-33016PF cells. The probability of a residual cell being present in a dose of vaccine is approximately 1 in 10(34). Residual MDCK-DNA is < or =10 ng per dose and the ss-propiolactone used to inactivate influenza virus results in reduction of detectable DNA to less than 200 base pairs (bp). Degenerate PCR and specific PCR confirm exclusion of oncogenic viruses. The manufacturing process has been validated for its capacity to remove and inactivate viruses. We conclude that the theoretical risks arising from manufacturing seasonal influenza vaccine using MDCK-33016PF cells are reduced to levels that are effectively zero by the multiple, orthogonal processes used during production.
基于细胞培养的生产方法可能有助于满足对季节性流感疫苗不断增长的需求，并发展应对流感大流行所需的生产灵活性。MDCK - 33016PF细胞用于生产基于细胞的季节性流感疫苗（Optaflu）；但是，与大多数连续细胞系一样，它能在免疫受损小鼠体内生长并产生肿瘤。因此，至关重要的是疫苗中不能残留细胞，细胞裂解物或DNA不具有致癌性，并且细胞基质不含有致癌病毒或致癌DNA。多个冗余过程确保了在MDCK - 33016PF细胞中生产的流感疫苗的安全性。一剂疫苗中存在残留细胞的概率约为10的34次方分之一。残留的MDCK - DNA每剂小于或等于10纳克，用于灭活流感病毒的β-丙内酯可使可检测到的DNA减少至小于200个碱基对（bp）。简并PCR和特异性PCR证实不存在致癌病毒。该生产工艺已验证其去除和灭活病毒的能力。我们得出结论，通过生产过程中使用的多个正交过程，使用MDCK - 33016PF细胞生产季节性流感疫苗所产生的理论风险已降低到有效为零的水平。
