METHOD OF PREPARING PROTECTIVE GLYCOPROTEIN OF MELIOIDOSIS PATHOGEN

摘要

FIELD: medicine, biochemistry. SUBSTANCE: invention relates to preparing glycoprotein antigen from B. pseudomallei. Method involves the use of two-day culture of virulent strain B. pseudomallei (Australian serovara) grown on cellophane film placed on solid nutrient medium surface. Culture is inactivated with acetone and an aqueous-salt extract is prepared by homogenization of 2% microbe cells suspension under mild conditions. Then homogeneity is subjected for successive fractionation by gel-filtration on carrier "Toyopearl HW-60", pH 7.2, and ion-exchange chromatography on DEAE-Sephadex A-50, pH 7.7, using 0.05 M tris-HCl as a buffer solution containing sodium chloride at concentration 0-1 M. Isolated glycoprotein consists of protein and carbohydrate at ratio 1:6 that shows homogeneity at immunoelectrophoresis assay. Significant protective effect is obtained after preliminary immunization of white mice with glycoprotein infected with virulent strains of Ps. pseudomallei. Preparation can be used for melioidosis vaccine making. EFFECT: improved method of preparing. 3 tbls