河豚毒素小分子拮抗肽的设计与鉴定

目的用计算机辅助分子设计技术,设计河豚毒素(TTX)小分子拮抗肽,分别用免疫学方法和动物实验进行验证。方法利用计算机辅助分子设计技术,对TTX的空间结构进行了优化;用分子对接确定了TTX关键位点;用分子模建设计了3个能与TTX结合的小分子拮抗肽;分别用竞争ELISA进行免疫学筛选;用动物实验进行中和活性测定。结果成功设计了3个针对TTX的小分子拮抗肽并进行了多肽合成,用竞争ELISA筛选到针对TTX的小分子拮抗肽P2,拮抗肽P2浓度与TTX的结合能力成正比。动物实验中,针对TTX的小分子拮抗肽P2对注射2.5倍半数致死剂量TTX昆明小鼠的保护率为25%,起到了一定解毒效果。结论获得了能与TTX特异性结合的中和性小分子拮抗肽,TTX浓度达到4.05μg/mL(LD50的2.5倍),TTX拮抗肽P2对实验组小鼠的保护率为25%。

Design and identification of antidote peptide against Tetrodotoxin

Objective: to design and synthesis antidote peptide of tetrodotoxin .assessment biological activity of antidote peptide with immunological assay and animal experiments.Methods: optimum spatial structure of TTX were obtained by computer aided molecular designing; critical site were determined by molecular butt joint.3 antidote peptides were designed and synthesized according to molecular simulation technique. biological activity binding with TTX were detected with antibody based competitive ELISA.neutralized capacity Were performed after hypoglossal vaccination Kunming mice .Results: 3 synthetic antidote peptides against TTX were obtained. a specific antidote peptide binding with TTX were selected by antibody based competitive ELISA. Binding capacity of P2 peptide had a direct proportion with concentration of P2 peptide. the neutralization antidote peptide With a Protection rate of 25% for Kunming mouse against 2.5 LD50 TTX attack. Conclusion: a neutralization antidote peptide were obtained with a Protection rate of 25 % for Kunming mouse against 2.5 LD50 TTX attack.