Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald - Insel Riems, Germany.
Vaccine. 2012 May 21;30(24):3603-9. doi: 10.1016/j.vaccine.2012.03.047. Epub 2012 Mar 31.
The World Health Organization (WHO) recommends the periodic evaluation of the purity of the cell lines used in the production of rabies vaccines, as well as the antigenic identity of the virus strains. Here, we analyzed seventeen marketed inactivated human rabies virus vaccines for viral and non-viral proteins by SDS-PAGE and Coomassie/silver staining. Mass spectrometric analysis of an abundant 60-70 kDa signal indicated that in most vaccines serum albumin of human origin (HSA) was the major component. Quantification of HSA in the vaccines revealed a mean concentration of 22 mg HSA/dose in all tested PVRV (purified vero cell rabies vaccine), HDCV (human diploid cell rabies vaccine) and PHK (primary hamster kidney) vaccines. In contrast, 1000-fold lower HSA levels and no HSA were detected in PCECV (purified chick embryo cell-culture vaccine) and PDEV (duck embryo rabies vaccine), respectively. Western blot analyses further confirmed a high bias in the HSA content, whereas the virus protein levels were rather similar in all tested vaccines. In addition, the vaccine viruses were sequenced within the N- and G-genes to identify the strain. In the majority of sequenced vaccines, the declared vaccine strain was confirmed. However, some discrepancies in the genetic identification were observed, supporting WHO's recommendation for the molecular characterization of vaccine seed strains. This research highlights the variation in purity found between different human rabies virus vaccines, and suggests that further research is needed to establish the impact non-active components have on the potency of such vaccines.
世界卫生组织（WHO）建议定期评估用于生产狂犬病疫苗的细胞系的纯度，以及病毒株的抗原身份。在这里，我们通过 SDS-PAGE 和考马斯亮蓝/银染色分析了 17 种市售的狂犬病病毒灭活疫苗中的病毒和非病毒蛋白。对丰富的 60-70 kDa 信号的质谱分析表明，在大多数疫苗中，人源血清白蛋白（HSA）是主要成分。对疫苗中 HSA 的定量分析显示，所有测试的 PVRV（纯化vero 细胞狂犬病疫苗）、HDCV（人二倍体细胞狂犬病疫苗）和 PHK（原代仓鼠肾）疫苗中 HSA 的平均浓度为 22 mg HSA/剂量。相比之下，在 PCECV（纯化鸡胚细胞培养疫苗）和 PDEV（鸭胚狂犬病疫苗）中，HSA 水平低 1000 倍，且未检测到 HSA。Western blot 分析进一步证实了 HSA 含量的高度偏差，而所有测试疫苗中的病毒蛋白水平则相当相似。此外，还对疫苗病毒进行了 N-和 G-基因测序，以鉴定毒株。在大多数测序疫苗中，均确认了所申报的疫苗株。然而，在遗传鉴定中观察到一些差异，支持了 WHO 对疫苗种子株进行分子特征分析的建议。本研究强调了不同狂犬病病毒疫苗之间发现的纯度差异，并表明需要进一步研究以确定非活性成分对这些疫苗效力的影响。
