【摘要】 目的制备α-芋螺毒素GI抗血清,为其中毒救治提供有效手段。方法构建包含α-芋螺毒素GI、PADRE和破伤风毒素C片段(TTC)融合蛋白的原核表达载体,原核表达GI-PADRE-TTC融合蛋白抗原,免疫BALB/c小鼠,获得抗血清。采用酶联免疫吸附实验(ELISA)检测血清抗体滴度,并测定其对小鼠的抗毒活性。结果获得GITTC融合蛋白69.5 mg。GI-TTC融合蛋白免疫小鼠第5次免疫后血清对GI、GI-BSA的抗体滴度分别达到1∶102 400及1∶204 800。100、200μl融合蛋白抗血清对2.0×LD50的α-芋螺毒素GI中毒小鼠存活率分别为14.3%、25.0%,且小鼠死亡时间显著延长。结论α-芋螺毒素GI与破伤风毒素C片段的融合蛋白具有较好的免疫原性,其产生的抗血清对α-芋螺毒素GI具有一定中和活性。更多还原

【Abstract】 Objective To prepare anti-serum of α-conotoxin GI and discover related treatments. Methods The genes of α-conotoxin GI,Pan HLA DR-binding epitope(PADRE)and Tetanus toxin fragment C(TTC)were constructed in the prokaryotic expression vector,and were expressed as fusion protein. Anti-serum was prepared with BALB/c mice immunized by GI-PADRE-TTC fusion protein. The titer of anti-serum and its antitoxic activity in mice were tested. Results69.5 mg of GI-PADRE-TTC fusion protein was prepared and the anti-serum titer was more than 1∶102 400(GI)and204 800(GI-BSA)after the fifth immunization in mice. The results of neutralization experiments showed that 14.3% and25.0% of the mice survived after the intraperitoneal injection of 100 and 200 μl anti-serum with 2.0×LD50 GI,respectively.The death time of mice was also prolonged significantly. Conclusion The fusion protein of α-conotoxin GI and Tetanus toxin fragment C possesses high immunogenicity and its anti-serum shows antitoxic activity.更多还原