purification & separation of bacillus anthracis 34f2 antigens with uf & hplc

anthrax is one of the important zoonose disease between human and cattle. bacillus anthracis is a gram- positive bacterium which its tripartite protein toxin contains protective antigen (pa), edema factor (ef) and lethal factor (lf) and a poly d- glutamic acid capsule. a recent resurgence of interest in b.anthracis resulted in improved methods for production, separation and purification of the known toxin components. this in turn has lead to development of much more sensitive toxin-detection systems. further understanding of toxin activity, the role of individual components in protection against the disease and the production of antigens for use in diagnostic requires high-degree of purity. in this study after cultivation of bacteria and separaing the supernatant the solution was passed through different membrane of  ultra filter system with 30000, 50000 and 100000 dalton molecular weight cut off respectively. the final solution was treated with ammonium sulphate and the antigens prepared which after dialysis have passed through gel chromatography of hplc for separating of  b.anthracis toxins. finaly three toxin antigens ef,lf and pa  with 86, 87 and 85 k dalton molecular weight of high purity were obtained.