Evaluation of a downstream process for the recovery and concentration of a Cell-Culture-Derived rVSV-Spike COVID-19 vaccine candidate

Autor: Meni Girshengorn, Ofir Israeli, Lilach Cherry, Edith Lupu, Arnon Tal, Rona Levy, Osnat Rosen, Arik Monash, Einat Toister, Yaron Kafri, Arik Makovitzki, Lilach Levin, Irit Simon, Eyal Dor, Ziv Oren, Elad Lerer, Idan Hefetz, Hanan Tzadok, Yaakov Adar, Eyal Epstein, Ophir Hazan
Rok vydání: 2021
Předmět:
PES
polyethersulfone
rVSV-S
recombinant vesicular stomatitis virus-ΔG-spike
COVID-19 Vaccines
MWCO
molecular weight cutoff
Coronavirus disease 2019 (COVID-19)
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
PFU
plaque-forming units
TMP
transmembrane pressure
ACE2
angiotensin-converting enzyme 2
DSP
downstream process
Article
MEM
minimal essential medium
HF-TFF
hollow fiber-tangential flow filtration
Downstream (manufacturing)
rVSV
SFM
serum-free medium
Animals
Humans
hc-DNA
host cell DNA
DO
dissolved oxygen
Pandemics
Infectivity
MOI
multiplicities of infection
Downstream process
General Veterinary
General Immunology and Microbiology
Chemistry
SARS-CoV-2
Public Health
Environmental and Occupational Health
Spike Protein
COVID-19
Endonuclease digestion
Antibodies
Neutralizing
Cell biology
Clarification
MT
multi-tray
Infectious Diseases
Recombinant viral vaccine
Cell culture
Spike Glycoprotein
Coronavirus
Nucleic acid
Molecular Medicine
Hollow fiber
HCP
host cell protein
NTU
nephelometric turbidity units
HR
horseradish peroxidase
Zdroj: Vaccine
ISSN: 1873-2518
Popis: rVSV-Spike (rVSV-S) is a recombinant viral vaccine candidate under development to control the COVID-19 pandemic and is currently in phase II clinical trials. rVSV-S induces neutralizing antibodies and protects against SARS-CoV-2 infection in animal models. Bringing rVSV-S to clinical trials required the development of a scalable downstream process for the production of rVSV-S that can meet regulatory guidelines. The objective of this study was the development of the first downstream unit operations for cell-culture-derived rVSV-S, namely, the removal of nucleic acid contamination, the clarification and concentration of viral harvested supernatant, and buffer exchange. Retaining the infectivity of the rVSV-S during the downstream process was challenged by the shear sensitivity of the enveloped rVSV-S and its membrane protruding spike protein. Through a series of screening experiments, we evaluated and established the required endonuclease treatment conditions, filter train composition, and hollow fiber-tangential flow filtration parameters to remove large particles, reduce the load of impurities, and concentrate and exchange the buffer while retaining rVSV-S infectivity. The combined effect of the first unit operations on viral recovery and the removal of critical impurities was examined during scale-up experiments. Overall, approximately 40% of viral recovery was obtained and the regulatory requirements of less than 10 ng host cell DNA per dose were met. However, while 86–97% of the host cell proteins were removed, the regulatory acceptable HCP levels were not achieved, requiring subsequent purification and polishing steps. The results we obtained during the scale-up experiments were similar to those obtained during the screening experiments, indicating the scalability of the process. The findings of this study set the foundation for the development of a complete downstream manufacturing process, requiring subsequent purification and polishing unit operations for clinical preparations of rVSV-S.
Databáze: OpenAIRE
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