title: Root cause investigation of a viral contamination incident occurred during master cell bank (MCB) testing and characterization--a case study.
abstract: An adventitious agent contamination occurred during a routine 9 CFR bovine viral screening test at BioReliance for an Eli Lilly Chinese Hamster Ovary (CHO) cell-derived Master Cell Bank (MCB) intended for biological production. Scientists from the sponsor (Eli Lilly and Company) and the testing service company (BioReliance) jointly conducted a systematic investigation in an attempt to determine the root cause of the contamination. Our investigation resulted in the identification of the viral nature of the contaminant. Subsequent experiments indicated that the viral contaminant was a non-enveloped and non-hemadsorbing virus. Transmission electron microscopy (TEM) revealed that the viral contaminant was 25-30 nm in size and morphologically resembled viruses of the family Picornaviridae. The contaminant virus was readily inactivated when exposed to acidic pH, suggesting that the viral contaminant was a member of rhinoviruses. Although incapable of infecting CHO cells, the viral contaminant replicated efficiently in Vero cell with a life cycle of approximately 16 h. Our investigation provided compelling data demonstrating that the viral contaminant did not originate from the MCB. Instead, it was introduced into the process during cell passaging and a possible entry point was proposed. We identified the viral contaminant as an equine rhinitis A virus using molecular cloning and DNA sequencing. Finally, our investigation led us to conclude that the source of the viral contaminant was the equine serum added to the cell growth medium in the 9 CFR bovine virus test.
authors.affiliation: Bioproduct Research and Development, Lilly Research Laboratories, Eli Lilly and Company, DC3941 Lilly Corporate Center, Indianapolis, IN 46285, USA. chen_dayue@lilly.com
authors.forename: Dayue
authors.initials: D
authors.lastname: Chen
authors.forename: Raymond
authors.initials: R
authors.lastname: Nims
authors.forename: Sandra
authors.initials: S
authors.lastname: Dusing
authors.forename: Pamela
authors.initials: P
authors.lastname: Miller
authors.forename: Wen
authors.initials: W
authors.lastname: Luo
authors.forename: Michelle
authors.initials: M
authors.lastname: Quertinmont
authors.forename: Bhavin
authors.initials: B
authors.lastname: Parekh
authors.forename: Josh
authors.initials: J
authors.lastname: Poorbaugh
authors.forename: Jeri Ann
authors.initials: JA
authors.lastname: Boose
authors.forename: E Morrey
authors.initials: EM
authors.lastname: Atkinson