流感裂解疫苗纯化及裂解工艺的优化

目的优化大规模生产的流感病毒裂解疫苗的纯化及裂解工艺。方法比较凝胶层析和蔗糖密度梯度超速离心的纯化效果,以及TritonX-100和去氧胆酸钠两种裂解剂在不同条件下的裂解效果。结果层析法纯度稍高于蔗糖密度梯度超速离心法,而蔗糖密度区带超速离心法的收率优于层析法。TritonX-100的裂解效果优于去氧胆酸钠,裂解剂浓度为0.5%或0.8%,裂解3或4h,裂解率达90%以上。疫苗在4℃保存1年后,所有检测项目均合格,4℃保存18个月和37℃保存28d后,HA含量仍保持在30μg/ml以上。结论已建立了切实可行的流感病毒裂解疫苗规模生产的纯化及裂解工艺。

Optimization of Procedure for Purification and Lysis of Influenza Virus Vaccine

Objective To optimize the procedure for purification and lysis of influenza virus vaccine.Methods Compare the purification efficacies by gel filtration chromatography and by sucrose density gradient centrifutgation,as well as the lysis efficacies with Triton X-100 and with sodium deoxycholate under various conditions.Results The purity of split influenza vaccine after purification by gel filtration chromatography was slightly higher than that by sucrose density gradient ultracentrifugation,while the recovery of vaccine by the latter was higher than that by the former.Triton X-100 showed good lysis efficacy as compared with sodium deoxycholate.After lysis with Triton X-100 at a final concentration of 0.5% or 0.8% for 3 or 4 h,more than 90% of influenza vaccine was lyzed.After storage at 4℃ for 12 months,all the quality indexes of the purified vaccine met the relevant requirements.However,after storage at 4℃for 18 months or at 37℃ for 28 d ,the HA content of vaccine was still more than 30 μg/ml.Conclusion A practical and feasible procedure for purification and lysis of influenza vaccine was developed.