Open AccessA simple novel approach for the purification of pertussis toxin
Author(s) -
Mégret Françoise,
Alouf Joseph E.
Publication year - 1988
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1988.tb02989.x
Subject(s) - bordetella pertussis , pertussis toxin , toxin , chinese hamster ovary cell , chromatography , bordetella , affinity chromatography , sepharose , polyacrylamide gel electrophoresis , chemistry , sodium dodecyl sulfate , gel electrophoresis , biochemistry , biology , microbiology and biotechnology , bacteria , g protein , receptor , enzyme , genetics
Pertussis toxin (PT) was purified from concentrated culture supernatants of Bordetella pertussis by a single step based on affinity chromatography on heparin‐Sepharose 6B with a recovery of 36% in two fractions. The fraction of highest specific activity (0.91 mg of toxin per mg protein) constituted 15.3% of toxin content in crude material and appeared homogeneous by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. It exhibited the five bands corresponding to toxin subunits. The purified fraction induced clustering of Chinese hamster ovary cells at as little as 0.06 ng per ml. PT solution gave a single precipitation line with rabbit immune serum raised against crude Bordetella pertussis supernatant.