冻干乙型脑炎减毒活疫苗非动物源性稳定剂的配方及其缓冲液的筛选

Screening of formula and buffer in non-animal-derived stabilizer of freeze-dried live attenuated Japanese encephalitis vaccine

目的筛选冻干乙型脑炎减毒活疫苗非动物源性稳定剂的配方及其缓冲液。方法分别以0.01 mol/L PBS和Earles液为缓冲液,右旋糖苷-70、蔗糖、山梨醇、乳糖及组氨酸不同组合构成的冻干稳定剂,制备乙型脑炎减毒活疫苗半成品及冻干疫苗成品,观察在不同温度存放不同时间的稳定性,并检测不同配方冻干稳定剂的崩解温度(Tc)。结果 12种配方冻干稳定剂制成的疫苗半成品可在4℃1周内保持滴度稳定性,在25℃3 d内保持滴度无明显变化,3 d后滴度大幅下降;以0.01 mol/L PBS为缓冲液的稳定剂制备的疫苗的Tc值低于Earles液,以Earles液为缓冲液的D2和F2配方制备的冻干疫苗成品,在4℃及37℃放置1周后,滴度均>5.7 logPFU/ml。结论已筛选出以Earles液为缓冲液,含右旋糖苷-70、蔗糖、山梨醇、乳糖及组氨酸的稳定剂配方,用于制备冻干乙型脑炎减毒活疫苗具有良好的保护效果。

Objective To screen the formula and buffer in non-animal-derived stabilizer of freeze-dried live attenuated Japanese encephalitis（JE） vaccine.Methods Final bulk and final product of freeze-dried live attenuated JE vaccine were prepared by various formula consisting of dextran-70,sucrose,sorbitol,lactose and histidine dissolved in 0.01 mol / L PBS and Earles solution respectively,and evaluated for stabilities after storage at various temperatures for various days.The disintegration temperatures（Tc） of the vaccine containing stabilizers of various formula were measured.Results The stabilities of final bulk of JE vaccine containing stabilizers of twelve formula were maintained after storage at 4℃ for one week.However,the titer of final bulk showed no significant change within 3 d after storage at 25 ℃,and decreased remarkably afterwards.The Tc of vaccine containing stabilizer using 0.01 mol / L PBS as buffer was lower than that using Earles solution.The titers of final product of freeze-dried live attenuated JE vaccines containing the stabilizers of formula D2 and F2 using Earles solution as buffer were more than 5.7 log PFU / ml after storage at 4 and 37 ℃ for one week.Conclusion The stabilizer consisting of dextran-70,sucrose,sorbitol,lactose and histidine using Earles solution as buffer was suitable for freeze-dried live attenuated JE vaccine.