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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。通过制备型SDS-PAGE和电渗方法从日本血吸虫成虫分离纯化出28KD,31/32KD及97KD蛋白。经BCA法检测蛋白含量,SDS-PAGE检测蛋白纯度和分子量,Western-blot及ELISA检测其抗原活性。结果日本血吸虫成虫28KD,31/32KD及97KD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;Western-blot均为单一反应区带;ELISA反应滴度分别为1∶640、1∶1280和1∶320。结果说明,由制备型SDS-PAGE和电渗析方法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,且不影响所纯化的抗原活性
In order to prepare sufficient and high purity polyvalent vaccine for the study of protective immunity against schistosomiasis. The 28KD, 31 / 32KD and 97KD proteins were isolated and purified from adult worms of Schistosoma japonicum by preparative SDS-PAGE and electroosmotic methods. The protein content was determined by BCA method. The purity and molecular weight of protein were detected by SDS-PAGE. The antigenic activity was detected by Western-blot and ELISA. Results The three purified proteins of 28KD, 31 / 32KD and 97KD from Schistosoma japonicum were single bands by SDS-PAGE and were consistent with the molecular weight of the isolated proteins. The protein contents were respectively 0.35mg / ml and 0.4mg / ml And 0.3mg / ml; Western-blot were single reaction zone; ELISA titers were 1: 640, 1:1280 and 1: 320 respectively. The results showed that the 28KD, 31 / 32KD and 97KD protein multivalent molecular vaccine of adult Schistosoma japonicum prepared by preparative SDS-PAGE and electrodialysis have the advantages of high purity, fast preparation, high recovery rate and the like without affecting the purified antigen active