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目的:为了制备足量、高纯度的供血吸虫保护性免疫力研究的多价分子疫苗。方法:通过制备型SDS-PAGE和电渗析方法从日本血吸虫成虫分离纯化出28 kD、31/32 kD及97kD蛋白。并检测其蛋白含量,蛋白纯度和分子量及其抗原活性。结果:此三种纯化蛋白经SDS-PAGE检测均为单一条带并与所分离的蛋白分子量相符;其蛋白含量分别为0.35mg/ml、0.4mg/ml和0.3mg/ml;Westem-blot均为单一反应区带;ELISA反应滴度分别为1:640、1:1280和1:320。结论:所制备的日本血吸虫成虫28kD、31/32kD及97kD蛋白多价分子疫苗,具有纯度高、制备快、回收率高等优点,而且不影响所纯化的抗原活性。
OBJECTIVE: To prepare a multivalent molecular vaccine for the study of protective immunity against Schistosoma japonicum in sufficient quantity and purity. METHODS: 28 kD, 31/32 kD and 97 kD proteins were isolated and purified from adult S. japonicum by preparative SDS-PAGE and electrodialysis. The protein content, protein purity, molecular weight and antigen activity were also tested. Results: The three purified proteins were single bands by SDS-PAGE and were consistent with the molecular weight of the isolated proteins. The protein contents were 0.35mg / ml, 0.4mg / ml and 0.3mg / ml respectively. Westem-blot As a single reaction zone; ELISA titers were 1: 640, 1: 1280 and 1: 320 respectively. CONCLUSION: The 28kD, 31 / 32kD and 97kD multivalent molecular vaccine prepared by adult Schistosoma japonicum have the advantages of high purity, fast preparation and high recovery rate, without affecting the purified antigen activity.