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将囊液抗原蛋白采用双向凝胶电泳分离,经考马斯亮蓝染色后用双向电泳图像分析软件分析蛋白点,以脑囊尾蚴病患者血清为一抗进行蛋白质印迹分析(Western blotting),选取阳性蛋白点进行质谱分析,获取肽质量指纹图谱,采用Mascot软件在NCBI中搜寻、比对,筛选出抗猪囊尾蚴IgG4特异性抗原。囊液抗原经双向电泳检测到(201±5)个蛋白斑点,相对分子质量(M_r)为10 000~170 000,等电点(pI)为3.0~10.0。Western blotting分析显示,与患者血清中特异IgG4抗体结合的抗原以小分子量蛋白为主,从中筛选出51个(抗原)点进行质谱分析,其中有21个点峰值大于39。对比分析显示,高峰值点与10种囊尾蚴蛋白有关,其中最为密切相关的为4种蛋白,按分值高低排序分别为Ts8B2、Ts8B3、10ku抗原和Sequence 3。
Cystic fluid antigen protein was separated by two-dimensional gel electrophoresis, stained with Coomassie brilliant blue after two-dimensional electrophoresis image analysis software analysis of protein spots in patients with cerebral cysticercosis serum as a primary antibody for Western blotting (Western blotting), select the positive protein Point mass spectrometry to obtain peptide mass fingerprinting, using Mascot software in NCBI search, comparison, screening anti-Cysticercus cellulosae IgG4-specific antigen. The cytoplasm antigen (201 ± 5) spots were detected by two-dimensional electrophoresis. The relative molecular mass (M_r) was 10 000 ~ 170 000 and the isoelectric point (pI) was 3.0 ~ 10.0. Western blotting analysis showed that 51 antigens were screened by mass spectrometry for the antigen binding to the specific IgG4 antibody in the sera of patients with small molecular weight proteins, of which 21 peaks were larger than 39. The comparative analysis showed that the peak value was related to 10 kinds of cyst protein, of which the four proteins were the most closely related, and were classified as Ts8B2, Ts8B3, 10ku antigen and Sequence 3 according to their scores.