[The isolation and purification of the protective antigen and the edema factor from the culture filtrate of Bacillus anthracis STI-1]. / Vydelenie i ochistka protektivnogo antigena i otechnogo faktora iz kul'tural'nogo fil'trata 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.

[Effect of anthrax toxin on the chemiluminescence of human leukocytes]. / Vliianie sibiriazvennogo toksina na khemiliuminestsentsiiu leikotsitov cheloveka.

The effect of substrate containing crude anthrax toxin on the phagocytosing leukocyte chemiluminescence has been studied. Preliminary toxin incubation with leukocytes for 60 min blocks cell chemiluminescence in the linear ratio effect concentration in the protein component of the toxin; the minimum concentration of the toxin protein component inhibiting the phagocytosing leukocyte luminescence is 3-5 micrograms per 5 x 10(5) cells. The substrate pure mixture of the oedema factor and protective antigen inhibits the chemiluminescence more intensively than crude toxin does. Inhibiting chemiluminescence activity of the anthrax toxin is directly proportional to its adenylate cyclase activity.

[The use of gel chromatography for the isolation of biologically active toxin complex from Bacillus anthrax]. / Primenenie gel'-khromatografii dlia vydeleniia biologicheski aktivnogo toksinnogo kompleksa sibireiazvennogo mikroba.

The multistep fractioning of the protein components from cultural filtrate of B. anthracis grown on casaminoacids containing medium was done. The scheme for preliminary purification of a toxin complex of B. anthracis against low and high molecular mass contaminants has been elaborated and includes ultrafiltration, gel chromatography in ultragel AcA-202 and TSK-gel toyopal HW-55. Biological activities of the complex,toxicity for laboratory animals and adenylate cyclase activity characteristic of B. anthracis toxin, are shown to remain intact in course of preliminary purification. Molecular masses of protein subunits from the fraction containing anthracis toxin activity reach 85-90 kD, 0.3-0.5 mg of toxin complex protein is yielded from 1 l of B. anthracis cultural filtrate.