SMITH DIFFUSE )TYP2( استخراج و خالص سازي كپسول استافيلوكوك آريوس

Extraction and purification of the capsule of Staphylococcus aureus type 2

History and Objectives: Extraction and purification of polysaccharide capsule is the first step for preparation preventive vaccines in infections as a result of encapsulated Staphylococcus aureus. This study was carried out to extract and purify Staphylococcus aureus type 2. Materials and Methods: The descriptive and explorative strategy of this study was conducted on Staphylococcus aureus (Smith diffuse variant) in BHI broth in a shaking incubator (200 rpm) in an aerobic conditions at a temperature of 37 °C. The capsule was extracted using dilute acid and heating method. Then, dialysis was done overnight at 4 °C and the product was lyophilized. Lyophilized powder was purified using a cellulose DEAE column (2.6 x 50 cm) by gradient method using bicarbonate ammonium buffers (0.01-1 M) at a velocity of 60 rat/It (Ion exchange chromatography). Finally, absorption spectrum was plotted and purified capsule was analyzed using scanning spectrophotometer. Results: Polysaccharide capsule was collected in final growth phase at 0.1 HCL and at a temperature of 100 °C. Absorption spectrum were evaluated using H2SO4/galacial acetic acid assay at wavelength range of 350-620 nm. The result showed that chromatographs are well matched with previously purified antigens. Conclusion and Recommendations: It is concluded that it is possible to extract and purify the capsule of Staphylococcus aureus type 2 and it is recommended to produce the related preventive vaccines.