title: Mycoplasma Clearance and Risk Analysis in a Model Bioprocess.
abstract: Mycoplasmas are a type of bacteria that lack cell walls and are occasional cell culture contaminants. In a biotechnology setting, because they can pass through 0.2 μm filters, mycoplasmas could pose a potential patient safety hazard if undetected contaminants from the production culture were not completely removed by downstream biotechnology manufacturing. In this study we investigated the ability of typical commercial monoclonal antibody purification operations to clear and kill mycoplasmas, usingAcholeplasma laidlawiias a model organism. Our spike/removal studies have shown that protein A column chromatography clears about 4-5 log10Column regeneration effectively preventsA. laidlawiicolumn carryover between chromatography runs. Moreover, low-pH hold steps, typically implemented after protein A purification, effectively killA. laidlawiiusing either pH 3.8 glycine or acetate solutions (LRV ≥5.30 and ≥4.57, respectively). Solvent/detergent treatment, used in some processes instead of low-pH hold, also completely kills highly concentratedA. laidlawii(LRV ≥5.95).LAY ABSTRACT:Biotechnology medicines need to be free from contaminating microorganisms such as mycoplasmas, a type of bacteria that can cause disease in humans (e.g., walking pneumonia). Here we show that some monoclonal antibody manufacturing steps can effectively clear and/or killAcholeplasma laidlawii, a model mycoplasma species used in our study. This provides an additional level of safety assurance of biotechnology medicines for patients.
authors.affiliation: Division II/Office of Biotechnology Products/Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20903. Views expressed in this article represent those of the authors and not necessarily policy or guidance from the U.S. Food and Drug Administration; and.
authors.forename: Julie
authors.initials: J
authors.lastname: Wang
authors.affiliation: Division II/Office of Biotechnology Products/Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20903. Views expressed in this article represent those of the authors and not necessarily policy or guidance from the U.S. Food and Drug Administration; and.
authors.forename: Sarah
authors.initials: S
authors.lastname: Johnson
authors.affiliation: Division II/Office of Biotechnology Products/Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20903. Views expressed in this article represent those of the authors and not necessarily policy or guidance from the U.S. Food and Drug Administration; and.
authors.forename: Matthew
authors.initials: M
authors.lastname: Brown
authors.affiliation: Division II/Office of Biotechnology Products/Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20903. Views expressed in this article represent those of the authors and not necessarily policy or guidance from the U.S. Food and Drug Administration; and.
authors.forename: Scott
authors.initials: S
authors.lastname: Lute
authors.affiliation: Division II/Office of Biotechnology Products/Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20903. Views expressed in this article represent those of the authors and not necessarily policy or guidance from the U.S. Food and Drug Administration; and.
authors.forename: Cyrus
authors.initials: C
authors.lastname: Agarabi
authors.affiliation: Division of Viral Products/Office of Vaccine Research and Review/Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20903.
authors.forename: Alena
authors.initials: A
authors.lastname: Dabrazhynetskaya
authors.affiliation: Division of Viral Products/Office of Vaccine Research and Review/Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20903.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Chizhikov
authors.affiliation: Division II/Office of Biotechnology Products/Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20903. Views expressed in this article represent those of the authors and not necessarily policy or guidance from the U.S. Food and Drug Administration; and Kurt.Brorson@fda.hhs.gov.
authors.forename: Kurt
authors.initials: K
authors.lastname: Brorson
mesh_terms: D000128:Acholeplasma laidlawii / Q000254:growth & development / Q000302:isolation & purification*; D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods / Q000592:standards*; D001709:Biotechnology / Q000379:methods / Q000592:standards*; D016466:CHO Cells; D003412:Cricetulus; D004340:Drug Contamination / Q000517:prevention & control*; D007700:Kinetics; D008962:Models, Theoretical; D009174:Mycoplasma / Q000254:growth & development / Q000302:isolation & purification*; D011786:Quality Control; D018570:Risk Assessment
keywords: Acholeplasma laidlawii; Bioprocessing; Cell culture; Low-pH hold; Mycoplasma; Protein A chromatography; Solvent/detergent; Spike/clearance