Use of a new RNA next generation sequencing approach for the specific detection of virus infection in cells

Brussel, Audrey; Brack, Kerstin; Muth, Erika; Zirwes, Rudolf; Cheval, Justine; Hébert, Charles; Charpin, J; Marinaci, Alice; Flan, Benoît; Ruppach, Horst; Beurdeley, Pascale; Éloit, Marc

doi:10.1016/j.biologicals.2019.03.008

Cited by 14 publications

(19 citation statements)

References 33 publications

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“…However, current HTS sample preparation approaches and bioinformatic pipelines are not as rapid as PCR and can take 7-10 days 58 . Additionally, determining if the contaminant identified in a nucleic acid-based assay is biologically active may require a different method, though we would note that HTS of viral RNA has been used to demonstrate a virus is biologically active 64 . Despite these challenges, testing should be conducted for ATMP production for samples taken before virus harvest (for viral-vectored gene therapy products) and at the end of the manufacturing process (for cell therapy products) so that if contamination with an adventitious virus is detected, informed decisions regarding product lot rejection can be made.…”

Section: Implications For Cell and Gene Therapy Manufacturersmentioning

confidence: 99%

Viral contamination in biologic manufacture and implications for emerging therapies

et al. 2020

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Section: Implications For Cell and Gene Therapy Manufacturersmentioning

confidence: 99%

Viral contamination in biologic manufacture and implications for emerging therapies

et al. 2020

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“… The development of five live, reference virus stocks for NGS evaluation and standardization and completion of 24-month stability studies [ 6 ]. The use of NGS for the detection of active viral RNAs by transcriptomics analysis [ 14 , 15 ]. …”

Section: Perspectives On Ngs For Adventitious Virus Detectionmentioning

confidence: 99%

“…Dr. Marc Eloit (Institut Pasteur and PathoQuest) described newly developed processes for specifically identifying cell infection by live viruses, either through the analysis of the cell transcriptome by stranded RNA analysis or by metabolic labelling of RNAs [ 14 , 15 ]. Tested cells can be master cell lines or cell therapy drug substances, or cells used in in vitro assays to amplify very small amounts of virus that are present in for instance working or master cell lines, in raw materials, or in seeds.…”

Section: Validation and Applications Of Ngs For Adventitious Virus Tementioning

confidence: 99%

“…The specificity of detection for infected cells using NGS coupled to metabolic labelling, limits time-consuming follow-up studies and adds mycoplasma detection and identification to virus testing. The limit of detection using EBV-infected B95-8 cells in a background of non-infected lymphoid cells has been evaluated [ 15 ]. In terms of the range of detection of RNA-Seq, adventitious virus has been identified in a human lymphoid cell line [ 15 ], and contamination by squirrel monkey retrovirus (SMRV) in a Vero cell line has also been identified [ 14 ].…”

Section: Validation and Applications Of Ngs For Adventitious Virus Tementioning

confidence: 99%

“…The limit of detection using EBV-infected B95-8 cells in a background of non-infected lymphoid cells has been evaluated [ 15 ]. In terms of the range of detection of RNA-Seq, adventitious virus has been identified in a human lymphoid cell line [ 15 ], and contamination by squirrel monkey retrovirus (SMRV) in a Vero cell line has also been identified [ 14 ]. This technique has also been demonstrated to enable the differentiation between active and inert sequences in persistent viral infections [ 14 ].…”

Section: Validation and Applications Of Ngs For Adventitious Virus Tementioning

confidence: 99%

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Report of the second international conference on next generation sequencing for adventitious virus detection in biologics for humans and animals

et al. 2020

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The IABS-EU, in association with PROVAXS and Ghent University, hosted the “2 nd Conference on Next Generation Sequencing (NGS) for Adventitious Virus Detection in Human and Veterinary Biologics” held on November 13 th and 14 th 2019, in Ghent, Belgium. The meeting brought together international experts from regulatory agencies, the biotherapeutics and biologics industries, contract research organizations, and academia, with the goal to develop a scientific consensus on the readiness of NGS for detecting adventitious viruses, and on the use of this technology to supplement or replace/substitute the currently used assays. Participants discussed the progress on the standardization and validation of the technical and bioinformatics steps in NGS for characterization and safety evaluation of biologics, including human and animal vaccines. It was concluded that NGS can be used for the detection of a broad range of viruses, including novel viruses, and therefore can complement, supplement or even replace some of the conventional adventitious virus detection assays. Furthermore, the development of reference viral standards, complete and correctly annotated viral databases, and protocols for the validation and follow-up investigations of NGS signals is necessary to enable broader use of NGS. An international collaborative effort, involving regulatory authorities, industry, academia, and other stakeholders is ongoing toward this goal.

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Reshaping cell line development and CMC strategy for fast responses to pandemic outbreak

Zhang

Wang

et al. 2021

Biotechnology Progress

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The global pandemic outbreak COVID‐19 (SARS‐COV‐2), has prompted many pharmaceutical companies to develop vaccines and therapeutic biologics for its prevention and treatment. Most of the therapeutic biologics are common human IgG antibodies, which were identified by next‐generation sequencing (NGS) with the B cells from the convalescent patients. To fight against pandemic outbreaks like COVID‐19, biologics development strategies need to be optimized to speed up the timeline. Since the advent of therapeutic biologics, strategies of transfection and cell line selection have been continuously improved for greater productivity and efficiency. NGS has also been implemented for accelerated cell bank testing. These recent advances enable us to rethink and reshape the chemistry, manufacturing, and controls (CMC) strategy in order to start supplying Good Manufacturing Practices (GMP) materials for clinical trials as soon as possible. We elucidated an accelerated CMC workflow for biologics, including using GMP‐compliant pool materials for phase I clinical trials, selecting the final clone with product quality similar to that of phase I materials for late‐stage development and commercial production.

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Use of a new RNA next generation sequencing approach for the specific detection of virus infection in cells

Cited by 14 publications

References 33 publications

Viral contamination in biologic manufacture and implications for emerging therapies

Viral contamination in biologic manufacture and implications for emerging therapies

Report of the second international conference on next generation sequencing for adventitious virus detection in biologics for humans and animals

Reshaping cell line development and CMC strategy for fast responses to pandemic outbreak

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