[The isolation and purification of the protective antigen and the edema factor from the culture filtrate of Bacillus anthracis STI-1].
A rapid method for producing a protective antigen (PA) and edema factor (EF), components of anthrax toxin, is described. The specific features of the method are as follows: addition of a mixture of protease inhibitors to the culture fluid; simultaneous concentration on a fiber filter and adsorption on hydroxylapatite, followed by non-linear gradient a phosphate concentration; purification of elution of a phosphate concentration; purification of eluates from salts via electrodialysis. Resultant from protein desorption, PA and EF agents are electrophoretically homogeneous by no less than 80%, 10 liters of filtrate yielded 8 mg of PA and 5 mg of EF. The resultant agents were tested for their biological properties and protective activity.
Rogov
S N
SN
Beznosov
M V
MV
Chugaĭ
V L
VL
Bashkova
V A
VA
D000818:Animals; D000881:Anthrax / Q000517:prevention & control; D000942:Antigens, Bacterial; D001408:Bacillus anthracis / Q000276:immunology* / Q000472:pathogenicity; D001427:Bacterial Toxins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification* / Q000633:toxicity; D003470:Culture Media; D004353:Drug Evaluation, Preclinical; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006168:Guinea Pigs; D007114:Immunization; D007123:Immunoelectrophoresis, Two-Dimensional / Q000379:methods; D051379:Mice; D008970:Molecular Weight; D051381:Rats; D011916:Rats, Inbred F344; D014462:Ultrafiltration / Q000295:instrumentation / Q000379:methods
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D003470:Culture Media; C030325:anthrax toxin