Open AccessAffinity purification of recombinant cholera toxin B subunit oligomer expressed in Bacillus brevis for potential human use as a mucosal adjuvant
Author(s) -
Yasuda Yoko,
Matano Keiko,
Asai Toru,
Tochikubo Kunio
Publication year - 1998
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.1998.tb01141.x
Subject(s) - cholera toxin , pentamer , recombinant dna , protein subunit , toxin , biology , affinity chromatography , microbiology and biotechnology , biochemistry , enzyme , gene
For use as a mucosal adjuvant for human vaccines, a simple method has been developed for the affinity purification of recombinant cholera toxin B subunit which had been expressed in a safe host, Bacillus brevis . Recombinant cholera toxin B subunit, adsorbed quantitatively to a d ‐galactose‐agarose column, was eluted with an 0.1–0.4 M d ‐galactose gradient with a yield of >90%. The cholera toxin B subunit preparation was similar to the native cholera toxin B subunit with respect to GM1 binding ability, remarkable stability of the pentamer, and the dissociation‐reassociation property by shifting pHs. Cross‐linking experiments with glutaraldehyde demonstrated that the pentameric form was predominant; tetrameric, trimeric, dimeric and monomeric forms were detected to a lesser extent, and additionally 10‐ and 15‐mers were observed depending on the concentration of the cholera toxin B subunit.