title: Case Study: A Novel Bacterial Contamination in Cell Culture Production--Leptospira licerasiae.
abstract: Leptospira licerasiae, a novel bacterial contaminant found in Genentech cell culture manufacturing operations, poses a challenge to current microbial control strategies in upstream cell culture processes, as this microorganism is fully capable of passing through 0.1 μm sterilizing-grade filtration and is not detectable by standard microbiological methods described in major pharmaceutical compendia for microbial screening and quantification required for release of raw materials, in-process intermediates, and finished products in biopharmaceutical production. The root cause investigation was greatly aided by the genetic identification of the contaminant and subsequent confirmation by cultural method and real-time polymerase chain reaction assay from the affected product batches. The purpose of this case study is to share knowledge on the novel contaminant, L. licerasiae, and potential routes of contamination in the cell culture manufacturing environment from a series of investigations involving root cause analysis, impact assessments, risk assessment, and global corrective and preventative action, as well as to provide guidance on the detection and prevention of Leptospira contamination with the intent to aid the industry to continually improve microbial control strategies for the benefit of patients.
  Leptospira licerasiae, a novel bacterial contaminant found in cell culture manufacturing operations, poses a challenge to current microbial control strategies in upstream cell culture processes because this microorganism is capable of passing through 0.1 μm sterilizing-grade membrane filters and is not detectable by standard microbiological methods used in biopharmaceutical production. The root cause investigation was greatly aided by the genetic identification of the contaminant and subsequent confirmation by cultural method and real-time polymerase chain reaction assay from the affected product batches. The purpose of this case study is to share knowledge on the novel contaminant, L. licerasiae, and potential routes of contamination in the cell culture manufacturing environment from a series of investigations to provide the industry with guidance on the detection and prevention of Leptospira contamination.
authors.affiliation: Genentech, Inc., San Francisco, CA.
authors.forename: Joseph
authors.initials: J
authors.lastname: Chen
authors.forename: Jesse
authors.initials: J
authors.lastname: Bergevin
authors.forename: Robert
authors.initials: R
authors.lastname: Kiss
authors.forename: Gordon
authors.initials: G
authors.lastname: Walker
authors.forename: Todd
authors.initials: T
authors.lastname: Battistoni
authors.forename: Patricia
authors.initials: P
authors.lastname: Lufburrow
authors.forename: Harry
authors.initials: H
authors.lastname: Lam
authors.forename: Anders
authors.initials: A
authors.lastname: Vinther
mesh_terms: D018929:Cell Culture Techniques; D015169:Colony Count, Microbial; D004340:Drug Contamination / Q000517:prevention & control; D004866:Equipment Contamination; D006801:Humans; D007919:Leptospira / Q000235:genetics; D008828:Microbiological Techniques; D060888:Real-Time Polymerase Chain Reaction; D018570:Risk Assessment