A new purification strategy for fraction 1 capsular antigen and its efficacy against Yersinia pestis virulent strain challenge.
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.
Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing 100071, China.
Tang
T
Wang
Zhizhen
Z
Qi
Benchuan
B
Wu
Ziwen
Z
Zhu
Yonghai
Y
Yang
Baizhong
B
Cui
Ruixia
R
Dai
Qingwen
Q
Zhang
Yefeng
Y
Qiu
Zuyun
Z
Wang
Hu
H
Wang
Zhaobiao
Z
Guo
Xiaoyi
X
Wang
Ruifu
R
Yang
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D014774:Virulence; D015010:Yersinia pestis / Q000276:immunology / Q000472:pathogenicity*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial