摘要
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。
The prepared cyst fluid antigen was separated by two-dimensional electrophoresis （2-DE）. The protein spots were analyzed by ImageMaster 2D Plantinum 6.0 software after Coomassie blue staining, on which Western blotting analysis was performed using sera of patients with cerebral eysticercosis as the primary antibody. The positive protein spots were selected and analyzed by Matrix-assisted laser desorption ionization tandem mass spectrometry （GC-MS）, and the peptide mass fingerprints were obtained. The specific Cysticercus cellulosae antigen for IgG4 antibody was identified with the Mascot software by searching and alignment in NCBI. 2-DE detected 201± 5 protein spots in the cystic acid antigen, with a relative molecular weight （Mr） of 10 000-170 000 and a pI of 3.0- 10.0. Western blotting analysis showed that the antigens binding the specific IgG4 antibody were mainly small molecular-weight proteins. Fifty-one （antigen） spots were obtained from them for mass spectrometry, and 21 of them were positive （peak score〉39）. The positive spots were related with 10 eysticereus proteins, most closely with Ts8B2, TsSB3, 10ku antigen and Sequence 3.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2017年第5期509-511,共3页
Chinese Journal of Parasitology and Parasitic Diseases
基金
山东省自然科学基金(No.ZR2015YL033
ZR2015YL051)
国家自然科学基金(No.30872208)
山东省医学科学院医药卫生科技创新工程~~
关键词
猪囊尾蚴
囊液抗原
IGG4
双向电泳
Cysticercus cellulosae
Cysticercus fluid antigens
IgG4
Two-dimensional electrophoresis
