Mondal S K, Neelima M, Seetha Rama Reddy K, Ananda Rao K, Srinivasan V A
Indian Immunologicals Limited, Gachibowli, Hyderabad 500 019, India.
Biologicals. 2005 Sep;33(3):185-9. doi: 10.1016/j.biologicals.2005.05.003.
The rabies vaccine is produced by inactivation of rabies virus propagated on BHK21 cells. In the rabies inactivation process, BEI is added at a final concentration of 1.6 mM to the viral harvest at 37 degrees C, followed by a second dose of BEI at 24 h post-inactivation. Inactivation was confirmed by the mice innocuity test and tissue culture amplification test as per B.P (Vet) 2004. Validation of test procedure is essential as per cGMP requirement. The dose of BEI was validated by using lower and higher concentrations of BEI in inactivation process. The study indicated that BEI at a lower concentration (0.4 mM) was able to inactivate the rabies virus within 30 h and the routine concentration (1.6 mM) of BEI is effective in inactivating rabies virus within 18 h. The amplification test used for confirming the inactivation of the live virus was validated by spiking the sample with different dilutions of pretitrated live rabies virus. The test revealed that the amplification method is sensitive to detect live rabies virus if present in the inactivated sample. The validation of BEI as an inactivant and the amplification test are discussed.
狂犬病疫苗是通过对在BHK21细胞上繁殖的狂犬病病毒进行灭活来生产的。在狂犬病灭活过程中，在37摄氏度下向收获的病毒中加入终浓度为1.6 mM的β-丙内酯（BEI），灭活后24小时再加入一剂BEI。根据《英国药典（兽医）》2004版，通过小鼠无害试验和组织培养扩增试验确认灭活效果。按照药品生产质量管理规范（cGMP）要求，对试验程序进行验证至关重要。在灭活过程中使用较低和较高浓度的BEI对BEI的剂量进行了验证。研究表明，较低浓度（0.4 mM）的BEI能够在30小时内灭活狂犬病病毒，常规浓度（1.6 mM）的BEI能在18小时内有效灭活狂犬病病毒。用于确认活病毒灭活的扩增试验通过向样品中加入不同稀释度的预先滴定的活狂犬病病毒进行了验证。试验表明，如果灭活样品中存在活狂犬病病毒，该扩增方法对其检测很敏感。文中讨论了BEI作为灭活剂的验证以及扩增试验。
