Development of a Rabies Vaccine in Cell Culture for Veterinary Use in the Lyophilized Form

Severo, Matheus Gonçalves

doi:10.1016/b978-0-444-63660-7.00021-8

Cited by 5 publications

(5 citation statements)

References 36 publications

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“…The ice crystals sublimate during primary drying ( Tang and Pikal, 2004 ). The moisture content of the samples after primary drying appears to be as high as 7%–8% ( Severo et al, 2013 ). The remaining water during secondary drying is generally bound to proteins strongly.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Processing Characteristics of Freeze-Dried Pork Powder for Meat Emulsion Gel

Lee¹,

Choi²,

Jo³

et al. 2021

Food Sci Anim Resour

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Section: Resultsmentioning

confidence: 99%

“…The remaining water during secondary drying is generally bound to proteins strongly. After the removal of bound water, only 1%–2% of the moisture remained in the samples following secondary drying ( Severo et al, 2013 ). In this study, the moisture content of the freeze-dried powder was 1.34% ( Table 1 ).…”

Section: Resultsmentioning

confidence: 99%

Processing Characteristics of Freeze-Dried Pork Powder for Meat Emulsion Gel

Lee¹,

Choi²,

Jo³

et al. 2021

Food Sci Anim Resour

View full text Add to dashboard Cite

show abstract

“…0.7-mL vials for use in the freeze-dried process. The protocol (Table 1) was established with some modi cations related to lyophilization described by Severo et al [24]. Mean residual moisture obtained was given in Table 2 for the freeze-dried vaccine and the results were found according to WHO standards [25].…”

Section: Production Of Experimental Rabies Vaccinementioning

confidence: 99%

Pilot-Scale Production of Lyophilized Inactivated Rabies Vaccine Candidate in Vero Cells Under Fully Animal Component-Free Conditions Using Microcarrier Technology and Laboratory Animal Trials

Onen¹,

Bezawada²

2022

Preprint

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The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate recombinant trypsin, a soybean trypsin inhibitor (STI) should be added to the medium. A protocol was first tested in T-flasks and then passaged to 500 ml and 3 L spinner flasks. Cell detachment was completed in 10–12 min, and 0.4 g/l STI was added to a 3L spinner and cells were transferred into a 30 L stirred tank bioreactor. On day 5, the cell density had reached its maximum (around 1.8 ×106 cells/ml). Cell infection, at an MOI of 0.3 with serum-free medium conditions, yielded a maximal rabies virus titer of 1.82×107 FFU/mL at 5 days. All cell culture conditions and virus growing kinetics in serum-free media were investigated. In conclusion, Vero cells were grown on Cytodex 1 with serum-free media and a high amount of rabies virus was obtained. Immune response to inactivated rabies virus vaccine candidate was determined in mice challenge. Also, we evaluated inactive rabies vaccine candidate safety, and immunogenicity in mice, sheep, horses, and cattle. We found that no horses, sheep, or cattle were given vaccine IM at 3.2 IU/dose exhibited any clinical sign of disease and all developed high VNA titers (up to 10,03 IU/ml) by 3–4 WPI. After the accelerated stability studies, the lyophilized inactivated rabies vaccine candidate showed enough antigenic potency (2.6 IU/ml) in the mouse challenge test. Also, 18-month long-term stability studies showed enough immune response (1.93 IU/ml) on day 14. The activity of the vaccine candidate showed a good immune response and safety criteria that meet WHO requirements. This is the first pilot-scale mammalian cell-based viral rabies vaccine production study using microcarrier technology reported in Turkey.

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“…0.7-mL vials for use in the freeze-dried process. The protocol (Table 1) was established with some modifications related to lyophilization described by Severo et al [24]. Mean residual moisture obtained was given in Table 2 for the freeze-dried vaccine and the results were found according to WHO standards [25].…”

Section: Production Of Experimental Rabies Vaccinementioning

confidence: 99%

Pilot-Scale Production of Lyophilized Inactivated Rabies Vaccine Candidate in Vero Cells under Fully Animal Component-Free Conditions Using Microcarrier Technology and Laboratory Animal Trials

Onen¹,

Bezawada²

2022

JBiSE

View full text Add to dashboard Cite

The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate recombinant trypsin, a soybean trypsin inhibitor (STI) should be added to the medium. A protocol was first tested in T-flasks and then passaged to 500 mL and 3 L spinner flasks. Cell detachment was completed in 10 -12 min, and 0.4 g/L STI was added to a 3L spinner, and cells were transferred into a 30 L stirred tank bioreactor. On day 5, the cell density had reached its maximum (around 1.8 × 106 cells/mL). At an MOI of 0.3 with serum-free medium conditions, cell infection yielded a maximal rabies virus titer of 1.82 × 10 7 FFU/mL at 5 days. All cell culture conditions and virus growth kinetics in serum-free media were investigated. In conclusion, Vero cells were grown on Cytodex 1 with serum-free media and a high amount of rabies virus was obtained. A mouse challenge was used to determine the immune response to an inactivated rabies virus vaccine candidate. Also, we evaluated inactive rabies vaccine candidate safety, and immunogenicity in mice, sheep, horses, and cattle. We found that no horses, sheep, or cattle who were given vaccine IM at 3.2 IU/dose exhibited any clinical sign of disease and all developed high VNA titers (up to 10.03 IU/mL) by 3 -4 WPI. After the accelerated stability studies, the lyophilized inactivated rabies vaccine candidate showed enough antigenic potency (2.6 IU/mL) in the mouse challenge test. Also, 18-month long-term stability studies showed enough immune response (1.93 IU/mL) on day 14. The activity of the vaccine candidate showed a good immune response and safety criteria that meet WHO requirements. This is the first pi-

show abstract

Development of a Rabies Vaccine in Cell Culture for Veterinary Use in the Lyophilized Form

Cited by 5 publications

References 36 publications

Processing Characteristics of Freeze-Dried Pork Powder for Meat Emulsion Gel

Processing Characteristics of Freeze-Dried Pork Powder for Meat Emulsion Gel

Pilot-Scale Production of Lyophilized Inactivated Rabies Vaccine Candidate in Vero Cells Under Fully Animal Component-Free Conditions Using Microcarrier Technology and Laboratory Animal Trials

Pilot-Scale Production of Lyophilized Inactivated Rabies Vaccine Candidate in Vero Cells under Fully Animal Component-Free Conditions Using Microcarrier Technology and Laboratory Animal Trials

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