【摘要】 目的:为探索α芋螺毒素MI的解毒方法,研究MI分支肽抗原的合成方法、免疫原性及抗血清对MI的解毒活性。方法:采用固相多肽合成法合成含炔基MI线性肽及含叠氮赖氨酸分支肽,空气氧化得含炔基MI,通过铜[Cu(Ⅰ)]催化叠氮-炔杂环加成反应合成多分支肽。多分支肽4次免疫BALB/c小鼠制备抗血清,ELISA法测定血清抗体滴度,然后MI与抗血清混合注射KM小鼠,测定抗血清的保护效果。结果:合成了MI八分支肽,N端连接的MI八分支肽的免疫原性差,10、20μg/只剂量组第4次免疫后的血清抗体滴度分别为1∶400及1∶6400;C端连接的MI八分支肽的免疫原性较高,10、20μg/只剂量组第4次免疫后的血清抗体滴度分别为1∶3200及1∶25 600。各组200μL抗血清与20μg/kg MI混合后注射小鼠,小鼠均死亡,且死亡时间与对照组相比无显著差异。结论:C端连接的MI八分支肽比N端连接的八分肽免疫抗原性高,但两者的抗血清对MI无显著解毒活性。更多还原

【Abstract】 Objective: In order to obtain detoxification method of α-conotoxin MI, the synthetic method, immunogenicity of MI branched peptides antigens as well as the detoxification activities of antiserum to mice were studied. Methods: Alkynyl-containing MI linear peptides and azide-lysine branched peptide were synthesized by solid phase peptide synthesis method. The alkynyl-containing MI peptides were synthesized by air oxidation. The MI branched peptides were synthesized via a copper(I)-catalyzed azide-alkyne cycloaddition(CuAAc) reaction. After four immunization, the serum antibody titers of MI branched peptides were determined in the BALB/c mouse model by ELISA. MI was then mixed with antiserum and injected into KM mice to determine the protective activities of antiserum. Results: Two eight branched peptides of MI were successfully synthesized. The N-terminally linked eight branched MI exhibits poor immunogenicity. The serum antibody titer of 10 and 20 μg/mice is 1∶400 and 1∶6400 after the fourth immunization. In contrast, the C-terminally linked eight branched MI has higher immunogenicity, at the same dose, the serum antibody titer is 1∶3200 and 1∶25 600, respectively. However, the neutralizing activity of antiserum of eight branched MI antigens is low, all mice died after injection of 200 μL of antiserum and 20 μg/kg MI, and show no significant difference in death time compared with the control group. Conclusion:The immunogenicity of C-terminally linked eight branched MI is higher than that of N-terminally linked eight branched MI, but their antiserums have not significant protective activities.更多还原