Wang Tang, Qi Zhizhen, Wu Benchuan, Zhu Ziwen, Yang Yonghai, Cui Baizhong, Dai Ruixia, Zhang Qingwen, Qiu Yefeng, Wang Zuyun, Wang Hu, Guo Zhaobiao, Wang Xiaoyi, Yang Ruifu
Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing 100071, China.
Protein Expr Purif. 2008 Sep;61(1):7-12. doi: 10.1016/j.pep.2008.05.003. Epub 2008 May 16.
F1 antigen is an attractive candidate for the development of a subunit vaccine against plague. In previous study, the extraction of this antigen from Yersinia pestis is characterized by using organic solvents. In this work, a new purification strategy that produced high-purity F1 antigen from Y. pestis EV76 was developed by the substitution of physical disruption for organic solvent one, followed by a combination of ammonium sulfate fractionation and Sephacryl S-200HR column filtration chromatography. As revealed in this study, this purification procedure is simple and effective, and avoids potential adverse effect on the antigen by organic solvents. Highly purified F1 that adsorbed to 25% (v/v) Al(OH)3 adjuvant in phosphate-buffered saline (PBS) induced very high titers of antibody to F1 in BALB/c mice and protected them (100% survival) against subcutaneous challenge with 10(4) CFU of Y. pestis virulent strain 141.
F1抗原是开发抗鼠疫亚单位疫苗的一个有吸引力的候选物。在先前的研究中，从鼠疫耶尔森氏菌中提取这种抗原的方法是以使用有机溶剂为特征的。在这项工作中，通过用物理破碎代替有机溶剂提取，随后结合硫酸铵分级分离和Sephacryl S - 200HR柱过滤色谱法，开发了一种从鼠疫耶尔森氏菌EV76中生产高纯度F1抗原的新纯化策略。如本研究所示，该纯化程序简单有效，并且避免了有机溶剂对抗原的潜在不利影响。在磷酸盐缓冲盐水（PBS）中吸附到25%（v/v）氢氧化铝佐剂上的高度纯化的F1在BALB/c小鼠中诱导出非常高滴度的抗F1抗体，并保护它们（100%存活）免受10(4) CFU鼠疫耶尔森氏菌强毒株141的皮下攻击。
