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Cited by 32Rabies epidemiology and controlProtein purification and stabilityViral Infectious Diseases and Gene Expression in Insects

Purification of rabies virus produced in Vero cells grown in serum free medium

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Trabelsi, Khaled, Ben Zakour, Meriem, Kallel, Héla·2019·Vaccine
DOI 10.1016/j.vaccine.2019.06.072
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Abstract

Rabies is a viral zoonosis caused by negative-stranded RNA viruses of the Lyssavirus genus. It can affect all mammals including humans. Dogs are the main source of human rabies deaths, contributing up to 99% of all rabies transmissions to humans. Vaccination against rabies is still the sole efficient way to fight against the disease. Cell culture vaccines are recommended by World Health Organization (WHO) for pre and post exposure prophylaxis; among them Vero cell rabies vaccines which are used worldwide. In this work we studied the purification of inactivated rabies virus produced in Vero cells grown in animal component free conditions, using different methods. Cells were grown in VP-SFM medium in stirred bioreactor, then infected at an MOI of 0.05 with the LP2061 rabies virus strain. Collected harvests were purified by zonal centrifugation, and by chromatography supports, namely the Capto Core 700 and the monolithic CIM-QA column. Generated data were compared in terms of residual DNA level, host cell proteins (HCP) level and the overall recovery yield. Rabies virus purification using the monolithic column resulted in the highest antigen recovery yield, equal to 94%. Capto Core 700 showed a lower yield, about 84%; whereas the purification yield by zonal centrifugation was equal to 60%. In terms of host cell residual DNA removal, zonal centrifugation was the most efficient method; the removal yield was equal to 88.5%; elimination of host cell DNA was slightly lower when using the monolithic CIM-QA (equal to 73%). Whereas Capto Core 700 showed the lowest level (49.2%). Host cell protein removal varied between 92.6% for the monolithic column and 78.6% for the zonal centrifugation. Capto Core 700 eliminated 86.5% of HCP.

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  • Gel chromatography is a common purification method in human rabies vaccine production, but it has limited efficiency in DNA removal.

    “Currently, gel chromatography is a common purification method in human rabies vaccine production, but it has limited efficiency in DNA removal [25].”

    From Evaluation of the Efficacy of the Vaccine Production Process in Removing Residual Host Cell DNA from the Vero Cell Rabies Vaccine (2024) · cites this paper as [25]

    The Discussion (page unavailable) cites examples of Sepharose gel chromatography and gel filtration used to purify rabies virus, supporting their use in vaccine production. It does not establish that gel chromatography is common or that it has limited efficiency in removing DNA; its DNA-removal results concern other purification methods. (LLM, conf=0.82)

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Cited by

Sorted by citations

Authors
Trabelsi, K., Ben Zakour, M., & Kallel, H.
Affiliations
Institut Pasteur de Tunis (TN)
Corresponding author
Kallel, Héla
ORCID
Venue
Vaccine
Publisher
Elsevier BV
Published
2019-11
Volume
37
Issue
47
Pages
7052-7060
ISSN
0264-410X (ISSN-L)
DOI
10.1016/j.vaccine.2019.06.072
OpenAlex
W2959309080
Cited by
32
References
0
Citations by year
2026: 2 · 2025: 2 · 2024: 7 · 2023: 4 · 2022: 7 · 2021: 3 · 2020: 6 · 2019: 1
FWCI
2.38
Type
journal article
Language
en
Open access
Licence
Locations
  • Vaccine (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • HAL (submittedVersion, cc-by-nc-nd)
Topics
  • Life Sciences › Immunology and Microbiology › Virology › Rabies epidemiology and control
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
Keywords
rabies virus, inactivated rabies vaccine, Vero cells, serum-free medium, virus purification, zonal centrifugation, monolithic chromatography, host cell DNA removal, host cell protein removal, stirred tank reactor, cell culture vaccine
MeSH terms
Animals, Antibodies, Viral, Chlorocebus aethiops, Rabies, Rabies Vaccines, Rabies virus, Vaccination, Vero Cells, Virus Cultivation, Culture Media, Serum-Free, Cell Culture Techniques, Bioreactors, Virus Inactivation

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