Effect of relevant culture parameters on Pertussis Toxin expression by Bordetella pertussis.
Whooping cough vaccines are produced using different ranges of cultivation conditions and medium compositions, which are known to influence growth rate, virulence factor production and degradation, as well as the virulence factors' association to the cell. This study quantifies the impact of individual parameters on Pertussis Toxin (PT) production, using an optimized chemically defined medium as starting point, rather than a complex medium. A number of chemicals that are identified affect both growth rate and virulence factor production, which occur at similar levels in various commonly used production media. Also, degradation by proteolytic activity is shown to be an important parameter to monitor, since it significantly affects the PT yield. Low sodium concentrations, i.e. 50-75 mM rather than the conventional 100-140 mM, significantly increase the growth rate of the organism, the final optical density, as well as the association of PT to the cells. The absolute amount of biomass produced measured as dry weight, is similar for all sodium concentrations tested, contrary to earlier work. While it is known that high iron concentrations inhibit virulence factor production, it is shown here that iron-limited growth results in very high specific PT production. This finding may be used to produce a whole-cell vaccine with little biomass per dose, reducing whole-cell vaccine toxicity. The Bordetella pertussis strain 509 used here produces 30% more PT at 34 than at 37 degrees C, a commonly used cultivation temperature. The data in this study show that existing production processes for cellular and acellular vaccines can in principle be optimised considerably by taking simple measures.
Synco Bio-Partners, Paasheuvelweg 30, 1105 BJ Amsterdam ZO, The Netherlands. marcel_thalen@synco-biopartners.com
Marcel
M
Thalen
Marian
M
Venema
Jan
J
van den IJssel
Luc
L
Berwald
Coen
C
Beuvery
Dirk
D
Martens
Johannes
J
Tramper
D001886:Bordetella pertussis / Q000187:drug effects* / Q000254:growth & development* / Q000378:metabolism; D002244:Carbon / Q000032:analysis; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D046508:Culture Techniques; D005978:Glutathione / Q000032:analysis / Q000494:pharmacology; D007501:Iron / Q000032:analysis / Q000494:pharmacology; D009525:Niacin / Q000032:analysis / Q000494:pharmacology; D009584:Nitrogen / Q000032:analysis; D037342:Pertussis Toxin / Q000096:biosynthesis*; D010567:Pertussis Vaccine / Q000096:biosynthesis*; D012964:Sodium / Q000032:analysis / Q000494:pharmacology
D003470:Culture Media; D010567:Pertussis Vaccine; D009525:Niacin; D002244:Carbon; D012964:Sodium; D007501:Iron; D037342:Pertussis Toxin; D005978:Glutathione; D009584:Nitrogen