无明胶保护剂在麻疹风疹联合减毒活疫苗中的应用

Application of gelatin-free stabilizer to live attenuated measles and rubella combined vaccine

目的研制一种安全有效的无明胶保护剂,并应用于麻疹风疹联合减毒活疫苗的制备。方法制备麻疹风疹联合减毒活疫苗原液,取同一批麻疹病毒原液,分别加入不同成分和配比的无明胶保护剂,制成成品,以含明胶保护剂作对照,检测成品外观、病毒滴度及水分,确定保护剂的成分,再确定保护剂的配比。将麻疹与风疹病毒原液混合,加入筛选出的无明胶保护剂,制备麻疹风疹联合减毒活疫苗(共3批),同时制备3批含明胶保护剂对照疫苗。按《中国药典》三部(2010版)要求对联合疫苗原液、半成品、成品进行检定;分别将制备的联合疫苗于2～8℃放置3和6个月,检测疫苗外观、病毒滴度及水分的变化,于37℃放置1、2、3、4周,检测病毒滴度的变化;按《中国药典》二部(2010版)要求进行过敏试验。结果含海藻糖和右旋糖酐成分的保护剂为首选保护剂。制备的无明胶保护剂联合疫苗原液、半成品及成品的各项检定指标均符合《中国药典》三部(2010版)要求;无明胶保护剂联合疫苗成品于2～8℃放置6个月的水分和病毒滴度及37℃放置4周的病毒滴度与对照组疫苗相比,差异均无统计学意义(P>0.05);注射了无明胶保护剂联合疫苗的豚鼠在试验期内均未发生过敏反应,符合《中国药典》二部(2010版)的要求。结论无明胶保护剂对麻疹和风疹病毒具有较好的保护作用。

Objective To prepare a safe and effective gelatin-free stabilizer and apply to live attenuated measles and rubella combined vaccine. Methods Bulk of live attenuated measles and rubella combined vaccine was prepared. The bulks of measles virus of the same batch were added with gelatin-free stabilizers with various formula respectively to prepare final product, then ob- served for appearance and determined for virus titer and moisture, using those prepared with gelatin-containing stabilizers as control, based on which the formula was optimized. The bulks of measles and rubella viruses were mixed and added with the optimized gelatin- free stabilizer to prepare three batches of live attenuated combined vaccine. However, three batches of vaccine were prepared with gelatin-containing stabilizer and served as control. Control tests were performed on the bulk, final bulk and final product of the com- bined vaccine according to the requirements in Chinese Pharmacopeia （Volume III, 2010 edition）. The prepared combined vaccine was stored at 2 -8 ℃ for 3 and 6 months respectively then observed for appearance and determined for virus titer and moisture, or at 37 ℃ for 1, 2, 3 and 4 weeks and determined for virus titer. Hypersensitive test was performed according to the requirements in Chinese Pharmacopeia （Volume Ⅱ, 2010 edition）. Results The optimal stabilizer consisted of trehalose and dextran. All the quali- ty indexes of prepared bulk, final bulk and final product of combined vaccine with gelatin-free stabilizer met the requirements in Chi- nese Pharmacopeia （Volume Ⅲ , 2010 edition）. The virus titer and moisture of final product of combined vaccine prepared with gelatin-free stabilizer after storage at 2 - 8 ℃ for 6 months as well as the virus titer after storage at 37 ℃ for 4 weeks showed no sig- nificant difference with those of control vaccine （P 〉 0. 05）. No hypersensitive reaction was observed in the guinea pigs immunized with the combined vaccine prepared with gelatin-free stabilizer, which met requirements in Chinese Pharmacopeia （Volume Ⅱ, 2010 edition）. Conclusion Gelatin-free stabilizer showed good protective effect on measles and rubella viruses.