Potential Vaccine for Anthrax
Annual rept. 1 Jun 1988-30 Sep 1989
LOUISVILLE UNIV KY
Pagination or Media Count:
The main aim of this study has been to isolate, purify, and resolve the structure of the cell wall polysaccharide pCHO of Bacillus anthracis, and to study its properties and distribution on the cell as well as spore surfaces. The distributions and stability of PA protective antigen and EA surface extractable antigen were also studied using various methods. In order to obtain the pCHO, B. anthracis Delta-Sterne cells were grown overnight in Penassay broth and disrupted by sonication. The cell wall preparation was purified by sequential extraction with 1 hot SDS ad several washes in water. Freeze dried cell wall 100 mg was treated with 20 ml 50 HF at 4 C for 18 hr. The preparation was centrifuged and the supernatant was added to cold absolute ethanol in 15 ratio and allowed to stand 30 min at 4 C. The precipitate that was formed pCHO was separated by centrifugation and washed 3 times with absolute ethanol. The pCHO was then dissolved in a ml of d-H2O and freeze-dried for future analysis.