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目的探索华支睾吸虫ESPs、虫卵和虫体水溶性组分中生物大分子极性和抗原性的差异并寻找可行的分离方法。方法用HPTLC法分离华支睾吸虫ESPs、虫卵和虫体的水溶性组分,同时结合免疫染色法研究极性差异组分的抗原性。结果用有机溶剂组合可将ESPs、虫卵和虫体水溶性组分中极性强弱差异的化合物分离开。氯仿︰甲醇︰水(11︰9︰2)溶剂组合对弱极性化合物各组分有较好的分离效果。与虫卵相比较,ESPs和虫体的弱极性组分较复杂。不能被有机溶剂展开的强极性组分的抗原性较强,未检测出弱极性组分的抗原性反应。通过有机溶剂提取的弱极性组分可完全在水中复溶并被有机溶剂重新展开。结论华支睾吸虫来源的水溶性组分中的弱极性化合物成分能通过有机溶剂组合进行有效分离。这为分离华支睾吸虫来源的脂类等弱极性生物大分子以及研究其病原相关分子模式的性质奠定了基础。
Objective To explore the differences of polarity and antigenicity of biomacromolecules in the water-soluble components of ESPs, eggs and worms of Clonorchis sinensis and to find a feasible separation method. Methods The water-soluble components of ESPs, eggs and worms of Clonorchis sinensis were isolated by HPTLC, and the antigenicity of the polar components were studied by immunostaining. Results The compounds with different polarities were separated from the water-soluble components of ESPs, eggs and worms by organic solvent combination. Chloroform: methanol: water (11: 9︰2) solvent combination of the components of the weak polar compounds have better separation. The weak polar components of ESPs and parasites are more complicated than the eggs. The strong polar component that can not be spread by the organic solvent has strong antigenicity, and the antigenic reaction of the weak polar component is not detected. The weakly polar components extracted by the organic solvent can be completely reconstituted in water and re-spread by the organic solvent. Conclusion The weak polar compounds in the water-soluble components of Clonorchis sinensis can be effectively separated by the combination of organic solvents. This laid the foundation for the isolation of weakly polar biomacromolecules such as lipids from Clonorchis sinensis and the study of their pathogen-related molecular patterns.