重组乙型肝炎疫苗工艺中去除甲醛添加的探讨

Elimination of formaldehyde addition in processing of recombinant hepatitis B vaccine

目的通过比较甲醛处理前后重组乙型肝炎病毒表面抗原(HBsAg)和疫苗(汉逊酵母)成品各项检测指标及稳定性的变化,评估甲醛在重组乙型肝炎疫苗工艺中的作用,为重组乙型肝炎疫苗工艺中是否可取消甲醛处理步骤提供参考。方法采用扫描电镜观察、HPLC检测和疫苗体内效力评价指标半数有效剂量(ED_(50))、乙肝疫苗常规检测项目比较甲醛处理前后HBsAg及疫苗成品的变化情况,通过稳定性考察确定甲醛对疫苗稳定性和有效期的影响。结果 HBsAg经甲醛处理后产生轻微聚合,但疫苗成品ED_(50)和其他各项检测指标均无明显变化,动物异常毒性合格。甲醛处理和未处理疫苗成品的稳定性考察结果相同。结论重组乙型肝炎疫苗生产工艺中甲醛处理步骤对疫苗成品检测指标及稳定性无影响,可考虑取消,从而提高乙肝疫苗的质量和安全性。

Objective To evaluate the role of formaldehyde in processing of hepatitis B（HepB）vaccine by comparison of various indexes and stabilities of HBsAg and final product of recombinant HepB vaccine prepared with Hansenula polymorpha before and after treatment with formaldehyde, and provide a reference for elimination of formaldehyde addition in processing of the vaccine. Methods Various indexes of HBsAg and final product of recombinant HepB vaccine before and after treatment with formaldehyde were compared by scanning electron microscopy, HPLC, in vivo potency test as well as routine control tests, while the effects of formaldehyde on stability and validity period were evaluated by stability test.Results After treatment with formaldehyde, HBsAg was slightly aggregated, while the final product showed no significant change in ED（50） or other indexes, and was qualified in abnormal toxicity test. However, the stabilities of final products before and after treatment with formaldehyde showed no significant difference. Conclusion The formaldehyde treatment in processing of recombinant HepB vaccine showed no effect on the quality indexes and stability of final product of vaccine, which might be eliminated to improved the quality and safety of vaccine.