title: Ensuring the safety of vaccine cell substrates by massively parallel sequencing of the transcriptome.
abstract: Massively parallel, deep, sequencing of the transcriptome coupled with algorithmic analysis to identify adventitious agents (MP-Seq™) is an important adjunct in ensuring the safety of cells used in vaccine production. Such cells may harbour novel viruses whose sequences are unknown or latent viruses that are only expressed following stress to the cells. MP-Seq is an unbiased and comprehensive method to identify such viruses and other adventitious agents without prior knowledge of the nature of those agents. Here we demonstrate its utility as part of an integrated approach to identify and characterise potential contaminants within commonly used virus and vaccine production cell lines. Through this analysis, in combination with more traditional approaches, we have excluded the presence of porcine circoviruses in the ATCC Vero cell bank (CCL-81), however, we found that a full length betaretrovirus related to SRV can be expressed in these cells, a factor that may be of importance in the production of certain vaccines. Similarly, insect cells are proving to be valuable for the production of virus like particles and sub-unit vaccines, but they can harbour a range of latent viruses. We show that following MP-Seq of the Trichoplusia ni (High Five cell line) transcriptome we were able to detect a contaminating, latent nodavirus and identify an expressed errantivirus genome. Collectively, these studies have reinforced the role of MP-Seq as an integral tool for the identification of contaminating agents in vaccine cell substrates.
authors.affiliation: BioReliance Corporation, 14920 Broschart Road, Rockville, MD 20850-3349, USA. david.onions@bioreliance.com
authors.forename: D
authors.initials: D
authors.lastname: Onions
authors.forename: C
authors.initials: C
authors.lastname: Côté
authors.forename: B
authors.initials: B
authors.lastname: Love
authors.forename: B
authors.initials: B
authors.lastname: Toms
authors.forename: S
authors.initials: S
authors.lastname: Koduri
authors.forename: A
authors.initials: A
authors.lastname: Armstrong
authors.forename: A
authors.initials: A
authors.lastname: Chang
authors.forename: J
authors.initials: J
authors.lastname: Kolman
mesh_terms: D000818:Animals; D030121:Betaretrovirus / Q000302:isolation & purification; D018929:Cell Culture Techniques / Q000592:standards; D002460:Cell Line; D002522:Chlorocebus aethiops; D004340:Drug Contamination / Q000517:prevention & control*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D007915:Lepidoptera; D030381:Nodaviridae / Q000302:isolation & purification; D013678:Technology, Pharmaceutical / Q000592:standards*; D059467:Transcriptome; D014612:Vaccines / Q000096:biosynthesis*
chemical_list: D014612:Vaccines