title: Properties of pertussis toxin B oligomer assembled in vitro from recombinant polypeptides produced by Escherichia coli.
abstract: The subunits that make up the pentameric B oligomer of pertussis toxin (S2, S3, S4, and S5) were individually synthesized as recombinant polypeptides in Escherichia coli, isolated as insoluble inclusion bodies, and assembled into a multimeric form in vitro by spontaneous association following treatment with a chaotropic agent, reduction, and reoxidation. The recombinant B multimer, purified by fetuin-Sepharose affinity chromatography, contained all four of the individual subunits and possessed the mitogenic and hemagglutinating activities characteristic of the native B oligomer. Immunization of mice with the recombinant B oligomer elicited antibodies that neutralized pertussis toxin in vitro and, moreover, provided protection in vivo against the leukocytosis-promoting activity of the toxin. These results demonstrate the potential for assembly of complex multimeric proteins from recombinant DNA-derived polypeptides and provide a novel means for production of an acellular pertussis vaccine component.
authors.affiliation: Amgen Inc., Amgen Center, Thousand Oaks, California 91320-1789.
authors.forename: W N
authors.initials: WN
authors.lastname: Burnette
authors.forename: J L
authors.initials: JL
authors.lastname: Arciniega
authors.forename: V L
authors.initials: VL
authors.lastname: Mar
authors.forename: D L
authors.initials: DL
authors.lastname: Burns
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis; D004926:Escherichia coli / Q000378:metabolism*; D007114:Immunization; D051379:Mice; D037342:Pertussis Toxin; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D010566:Virulence Factors, Bordetella / Q000096:biosynthesis / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin