毕赤酵母表达的含前S抗原重组乙型肝炎疫苗的稳定性

目的考察毕赤酵母表达的含前S抗原重组乙型肝炎疫苗原液和成品的稳定性。方法将3批疫苗原液置4℃存放,分别于0、1、2、3、6个月时取样,测定其在各时间点的乙型肝炎表面抗原S、前S1(PreS1)、前S2(PreS2)抗原滴度及HPLC纯度;将3批疫苗成品置25℃和4℃存放,进行加速稳定性和长期稳定性试验,分别于0、1、2、3、6个月和0、3、6、9、12、18、24个月时取样,进行小鼠效力、pH值、细菌内毒素和无菌测定。结果 3批疫苗原液于4℃保存6个月,表面抗原S、PreS1、PreS2抗原滴度及HPLC纯度均无明显变化;3批疫苗成品于25℃放置6个月和4℃放置24个月,各个时间点的小鼠效力均符合暂定规程的要求,pH值保持稳定,细菌内毒素和无菌试验均符合《中国药典》三部(2005和2010版)要求。结论毕赤酵母表达的含前S抗原重组乙型肝炎疫苗原液及成品稳定性良好。

Stability of recombinant hepatitis B vaccine containing PreS antigen prepared with Pichia pastoris

Object ive To investigate the stability of bulk and final product of recombinant hepatitis B(HB)vaccine containing PreS antigen prepared with Pichia pastoris.Methods Three batches of bulk were stored at 4 ℃,from which samples were taken 0,1,2,3 and 6 months after storage and determined for titers of HBsAg(S),PreS1 and PreS2 as well as the HPLC purity.Three consecutive batches of final products were stored at 25 and 4 ℃ for accelerated and long-term stability tests respectively.Samples were taken from the final products 0,1,2,3 and 6 months after storage at 25 ℃ and from those 0,3,6.9,12,18 and 24 months after storage at 4 ℃,and tested for potency in mice,pH value,endotoxin and sterility.Results No significant changes were observed in the titers of S,PreS1 and PreS2 antigens or HPLC purities of bulks after storage at 4 ℃ for 6 months.The potencies in mice of three batches of final products after storage at 25 ℃ for 6 months and at 4 ℃ for 24 months met the temporary requirements,while the pH values were stable,and the endotoxin contents and sterilities met the requirements in Chinese Pharmacopoeia(Volume Ⅲ,2010 edition).Conclusion The bulk and final product of recombinant hepatitis B vaccine containing PreS antigen prepared with P.pastoris showed high stability.