Extraction and purification of the capsule of Staphylococcus Aureus type 2
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Sayed asghar Havaie * , Taghi Naserpour Farivar , Mahmoud Saffari  |
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Abstract: (11239 Views) |
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 condition at a temperature of 37°C. The capsule was extracted using dilute acid and heating method. Then, dialysis was done overlight at 4°C and the product was lyophilized. Lyophilized powder was purified using a cellulose DEAE column (2.6×50 cm) by gradient method using bicarbonate ammonium buffers (0.01-1 M) at a velocity of 60 mt/h (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. |
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Keywords: Staphylococcus aureus, Polysaccharide capsule, Final growth phase, Purified antigen, Extraction |
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Full-Text [PDF 1047 kb]
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Type of Study: Research |
Subject:
General Received: 2008/11/10 | Published: 2001/10/15
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