Preparation and characterization of Pertussis toxin subunits.
Pertussis toxin (PT), a typical A-B oligomer exotoxin of Bordetella pertussis, has been demonstrated to be an essential protective antigen for acellular pertussis vaccine against whooping cough. In order to investigate the associated functionality ascribed to its components, we have purified A and B oligomers for the activity study. The A oligomer (S1 subunit) of PT was expressed in E. coli B834 (DE3) harboring expression vector (pET-20b) with the insert of S1 coding region and purified by metal-chelating column. The B oligomer was isolated by a single-step purification procedure. Individually, recombinant S1 and B oligomer exhibited quite distinct biological activities in vivo. S1 subunit induced leukocytosis-promoting (LP) activity, but did not affect mouse body weight-gain. On the contrary, B oligomer reduced mouse body weight-gain but did not reveal LP activity. In vitro, the combination of S1 subunit and B oligomer could enhance the toxic activities as exhibited by native PT and showed an additive toxicity in CHO cell clustering test and hemagglutination assay.
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Division of Biologics Quality Control, National Institute of Preventive Medicine, Taipei, Taiwan, ROC.
Wo
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Lu
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D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D005260:Female; D006384:Hemagglutination; D007964:Leukocytosis / Q000209:etiology; D051379:Mice; D008813:Mice, Inbred ICR; D037342:Pertussis Toxin; D011994:Recombinant Proteins / Q000633:toxicity; D010566:Virulence Factors, Bordetella / Q000737:chemistry* / Q000302:isolation & purification / Q000633:toxicity; D015430:Weight Gain / Q000187:drug effects
D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin