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Production and purification of recombinant protective antigen and protective efficacy against Bacillus anthracis
Author(s) -
Julie Ann Miller,
B.W. McBride,
R. J. Manchee,
Philippa Moore,
Les Baillie
Publication year - 1998
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.1998.00274.x
Subject(s) - bacillus anthracis , recombinant dna , microbiology and biotechnology , antigen , bacillaceae , bacillales , bacteria , biology , bacillus (shape) , virology , chemistry , immunology , biochemistry , gene , bacillus subtilis , genetics
Recombinant protective antigen (rPA), expressed by Bacillus subtilis WB600 (pPA101), has been purified to homogeneity and the protective efficacy against a Bacillus anthracis challenge has been investigated. rPA was fractionated from culture supernatant fluid by ammonium sulphate, followed by anion exchange chromatography using DEAE Streamline™, anion‐exchange chromatography on FPLC MonoQ HR 10/10 and finally, gel filtration chromatography on FPLC Superose 12 HR 10/30, to yield 7 mg rPA per litre of culture. The protective efficacy of rPA against an airborne challenge with the AMES strain of B. anthracis was determined in the presence of the adjuvants, alhydrogel and Ribi, and compared to that achieved by the current UK human vaccine in guinea pigs. rPA combined with the Ribi adjuvant was found to provide 100% protection against challenge.