Large-scale production of Vibrio cholerae toxin B subunit for use in oral vaccines.
By systematically manipulating promoter and ribosome binding structures, plasmid copy number and the structure of the cholera toxin B (CTB) subunit gene, we were able to develop a plasmid expression system that, when used in conjunction with an optimized growth medium, provided yields of CTB approaching one gram per liter. The CTB protein which was secreted to > 95%, could readily be purified from the growth medium of a V. cholerae production strain and was shown to be immunologically indistinguishable from previously used vaccine preparations of native or recombinant CTB.
Lebens
M
M
University of Göteborg, Dept. of Medical Microbiology and Immunology, Sweden.
Johansson
S
S
Osek
J
J
Lindblad
M
M
Holmgren
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002772:Cholera Toxin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D007114:Immunization; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D011817:Rabbits; D014614:Vaccines, Synthetic / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D014734:Vibrio cholerae / Q000235:genetics*
D014614:Vaccines, Synthetic; D002772:Cholera Toxin