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目的:初步分析由我国炭疽疫苗株A16R所生产的无菌培养滤液中的主要成分。方法:运用免疫蛋白质组学的手段,从炭疽A16R疫苗株无菌培养滤液的二维凝胶电泳中选择与保护性抗体结合和不与该血清结合但丰度高的蛋白质点,进行质谱鉴定和信号肽分析。结果:共选择73个点,从炭疽芽孢杆菌数据库中鉴定出66个。在66个点中,有43个点与保护性血清结合;23个为高丰度蛋白,不与该血清结合。在与保护性血清结合的43个点中,13个点为不同大小的PA分子,其中相对分子质量83×103和63×103占主导,相对分子质量较小的片段仅4个。其余成分包括炭疽表面蛋白EA1、Sap、胶原黏附蛋白、伴侣分子DnaK等。结论:我们制备的炭疽无菌培养滤液中含有PA成分,PA是炭疽疫苗的重要保护性抗原。但PA以外的其他成分在抗炭疽免疫中起何种作用,是否参与诱发机体免疫应答有待进一步研究。
OBJECTIVE: To analyze the main components in aseptic culture filtrate produced by anthrax vaccine strain A16R in China. Methods: By means of immuno proteomics, two-dimensional gel electrophoresis of anthrax A16R vaccine strain was used to identify the protein spots which were bound to the protective antibody but not to the serum with high abundance, Signal peptide analysis. Results: A total of 73 spots were selected and 66 were identified from the B. anthracis database. Of the 66 spots, 43 spots were bound to protective serum; 23 were highly abundant and did not bind to this serum. Among the 43 spots that bind to the protective serum, 13 spots were PA molecules with different sizes, of which 83 × 103 and 63 × 103 molecules dominated, while only 4 had relatively small molecular weight. The remaining ingredients include anthrax surface protein EA1, Sap, collagen adhesion protein, chaperones DnaK and so on. CONCLUSION: The anthrax culture filtrate we prepared contains PA component and PA is an important protective antigen for anthrax vaccine. However, what is the role of other components besides PA in anti-anthrax immunity? Whether to participate in the induction of immune response remains to be further studied.