重组相思子毒素B链蛋白冻干保存与免疫效果评价  

Study on the immunogenicity of lyophilized formulation of recombinant abrin B chain protein

作　　者：王俊虹[1,2] 魏茂提 武佳[1] WANG Jun-hong;WEI Mao-ti;WU Jia(Logistics University of PAPF , Tianjin 300309, Chin)

机构地区：[1]武警后勤学院,天津300309 [2]天津市职业与环境危害防制重点实验室

出　　处：《中国预防医学杂志》2017年第11期857-860,共4页Chinese Preventive Medicine

基　　金：武警后勤学院开放基金项目(WHKF201503);军事预防医学"2110"三期工程建设项目

摘　　要：目的探讨重组相思子毒素B链蛋白(rATB)的冻干保存条件及其免疫雌性Balb/C小鼠后的免疫效果评价。方法 rATB分别与不同冻干保护剂配方混合进行真空冷冻干燥,复融后免疫小鼠4次。每次免疫后,间接ELISA方法检测小鼠血清中IgG效价,四免后采用相思子毒素(abrin toxin,AT)攻毒,并进行小鼠体外中和试验。结果 rATB与10g/L的蔗糖混合保存所测浓度最高,纯度可达98%以上。四免后rATB组血清IgG效价与PBS对照组比较差异有统计学意义(P<0.05)。攻毒保护试验和体外中和试验结果说明,rATB可以抵抗5×LD50的天然AT攻毒。结论 rATB和10g/L蔗糖混合冻干条件下所测定的浓度最高,且冻干蛋白复融后免疫小鼠后显示出良好的免疫原性。Objective To evaluate the immunogenicity of lyophilized formulation of recombinant abrin B chain protein（rATB）. Methods rATB was mixed with sucrose at 10 g/L or 25 g/L concentration and with/without glycerol and then lyophilized.After reconstituted with PBS,rATB was used to vaccinate Balb/C mice for4 times.IgG antibody was detected by ELISA after each vaccination.After vaccination,mice were challenged with abrin toxin（AT）and then the protein neutralization assay was performed and efficacy of passive protection was evaluated. Results A lyophilized formulation of rATB mixed with 10 g/L sucrose was stable and the purity of the recombinant protein was over 98%.IgG antibody level was significantly higher in mice vaccinated with rATB than PBS control（P〈0.05）.Meanwhile,all Balb/C mice survived after being challenged with 5×LD50 of native AT. Conclusions The purity of lyophilized rATB with 10 g/L sucrose is the highest,which also triggers a good immune response and protective efficacy against exposure to AT.

关 键 词：相思子毒素 B链 疫苗 冻干 免疫效果 评价 

分 类 号：R392-33[医药卫生—免疫学]