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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 investigate the freeze-dried storage conditions of recombinant acitreotoxin B chain protein (rATB) and the immunological effects of immunized female Balb / C mice. Methods rATB were mixed with different lyoprotectant formulations were vacuum freeze-dried, mice were immunized 4 times after thawing. After each immunization, indirect ELISA was used to detect the IgG titer in the serum of mice. After four immunizations, the mice were challenged with abrin toxin (AT) and the mice were in vitro neutralized. Results rATB and 10g / L of sucrose mixed stored highest concentration, purity up to 98%. The serum IgG titer in rATB group after four-no-exemption was significantly lower than that in PBS control group (P <0.05). The challenge test and the in vitro neutralization test result indicate that rATB can resist 5 × LD50 natural AT challenge. Conclusion The concentration of rATB and 10g / L sucrose was the highest under the mixed freeze-drying conditions, and the immunized mice showed good immunogenicity after reconstitution of lyophilized protein.