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Evaluation of Pertussis Toxin Expression in B2 and THIJS Media
H Moradtalab1, M Noofeli2,2, H Zeighami1
1Department of Microbiology and Virology, Faculty of Medical Sciences, Zanjan University of Medical Sciences, Zanjan, Iran.
Insights
Thalen-IJssel (THIJS) medium enhances Bordetella pertussis growth and pertussis toxin (PT) production compared to B2 medium. This suggests THIJS medium could improve whole-cell pertussis vaccine (wP) production, potentially reducing vaccine toxicity.
Area of Science:
- Microbiology
- Vaccine Development
- Biotechnology
Background:
- Whole-cell pertussis vaccine (wP) is crucial for preventing pertussis, with pertussis toxin (PT) being a key virulence factor.
- Current wP production relies on bioreactor culture of B. pertussis in B2 medium.
- Optimizing culture media is essential for enhancing vaccine efficacy and safety.
Purpose of the Study:
- To evaluate the production of B. pertussis strain 509 PT in two different media: B2 and Thalen-IJssel (THIJS).
- To compare bacterial growth rates and PT yields using Chinese Hamster Ovary (CHO) cell and ELISA methods.
- To determine the optimal medium for increased PT production and potential reduction in vaccine toxicity.
Main Methods:
- Culturing B. pertussis strain 509 in both B2 and THIJS media under controlled bioreactor conditions.
- Monitoring bacterial growth rate via opacity measurements during the log growth phase.
- Quantifying cell-associated and released PT using ELISA and CHO cell assays at regular intervals.
Main Results:
- THIJS medium significantly increased the bacterial growth rate compared to B2 medium.
- Bacterial concentrations after 29 hours were higher in THIJS (29 IOU) than in B2 (19 IOU).
- THIJS medium resulted in higher cell-associated PT production (ODs of 1.6 and 1.1) compared to B2 medium (ODs of 1.1 and 0.9).
Conclusions:
- B. pertussis strain 509 cultured in THIJS medium yields higher cell mass and cell-associated PT than in B2 medium.
- THIJS medium is a promising alternative for wP production, potentially allowing for lower biomass per dose.
- Utilizing THIJS medium could lead to a more potent wP with reduced toxicity.
Abstract:
Whole-cell pertussis vaccine (wP) has been imperative and highly effective in preventing childhood deaths due to pertussis. Pertussis toxin is one of the virulence factors of Bordetella pertussis in all available pertussis vaccines. wP production in Razi Vaccine and Serum Research Institute is according to bioreactor culture of B. pertussis strains in B2 medium. The aim of this study was to evaluate B. pertussis strain 509 PT production in B2 and Thalen-IJssel (THIJS) media by Chinese Hamster Ovary (CHO) cell and enzyme-linked immunosorbent assay methods (ELISA). In the current study, B. pertussis strain 509 was cultured in B2 and THIJS media. Six samples were taken during the log growth phase within 2-3 h intervals (triplicate). The growth rate was calculated using opacity and the quantification of cell-associated and released PT measured by ELISA and CHO cell assays. THIJS medium was significantly showed an increase in the bacterial growth rate. During the first 29 h, bacterial concentrations in B2 and THIJS culture medium were 19 and 29 IOU, respectively. In THIJS medium, greater amount of pertussistoxin production was cell-associated. In B2 medium, maximum cell-associated toxin by ELISA and CHO cell assays were in the ODs of 1.1 and 0.9 and for THIJS medium in the ODs of 1.6 and 1.1, respectively. B. pertussis strain 509 in THIJS medium produced higher cell mass and cell-associated pertussis toxin than that of B2. It can be used for the production of whole-cell vaccine with higher pertussis toxin and accordingly using lower biomass per dose leading to the reduction of vaccine toxicity.
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