title: Extractables analysis of single-use flexible plastic biocontainers.
abstract: Studies of the extractable profiles of bioprocessing components have become an integral part of drug development efforts to minimize possible compromise in process performance, decrease in drug product quality, and potential safety risk to patients due to the possibility of small molecules leaching out from the components. In this study, an effective extraction solvent system was developed to evaluate the organic extractable profiles of single-use bioprocess equipment, which has been gaining increasing popularity in the biopharmaceutical industry because of the many advantages over the traditional stainless steel-based bioreactors and other fluid mixing and storage vessels. The chosen extraction conditions were intended to represent aggressive conditions relative to the application of single-use bags in biopharmaceutical manufacture, in which aqueous based systems are largely utilized. Those extraction conditions, along with a non-targeted analytical strategy, allowed for the generation and identification of an array of extractable compounds; a total of 53 organic compounds were identified from four types of commercially available single-use bags, the majority of which are degradation products of polymer additives. The success of this overall extractables analysis strategy was reflected partially by the effectiveness in the extraction and identification of a compound that was later found to be highly detrimental to mammalian cell growth.
  
  
  
  The usage of single-use bioreactors has been increasing in biopharmaceutical industry because of the appealing advantages that it promises regarding to the cleaning, sterilization, operational flexibility, and so on, during manufacturing of biologics. However, compared to its conventional counterparts based mainly on stainless steel, single-use bioreactors are more susceptible to potential problems associated with compound leaching into the bioprocessing fluid. As a result, extractable profiling of the single-use system has become essential in the qualification of such systems for its use in drug manufacturing. The aim of this study is to evaluate the effectiveness of an extraction solvent system developed to study the extraction profile of single-use bioreactors in which aqueous-based systems are largely used. The results showed that with a non-targeted analytical approach, the extraction solvent allowed the generation and identification of an array of extractable compounds from four commercially available single-use bioreactors. Most of extractables are degradation products of polymer additives, among which was a compound that was later found to be highly detrimental to mammalian cell growth.
authors.affiliation: Drug Product Development, Amgen Inc., Thousand Oaks, CA;
authors.forename: Liliana
authors.initials: L
authors.lastname: Marghitoiu
authors.affiliation: Drug Product Development, Amgen Inc., Thousand Oaks, CA;
authors.forename: Jian
authors.initials: J
authors.lastname: Liu
authors.affiliation: Drug Product Development, Amgen Inc., Thousand Oaks, CA;
authors.forename: Hans
authors.initials: H
authors.lastname: Lee
authors.affiliation: Drug Product Development, Amgen Inc., Thousand Oaks, CA;
authors.forename: Lourdes
authors.initials: L
authors.lastname: Perez
authors.affiliation: Drug Product Development, Amgen Inc., Thousand Oaks, CA;
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Fujimori
authors.affiliation: Drug Product Development, Amgen Inc., Thousand Oaks, CA;
authors.forename: Michael
authors.initials: M
authors.lastname: Ronk
authors.affiliation: Product Attribute Sciences, Amgen Inc., Thousand Oaks, CA;
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Hammond
authors.affiliation: Cell Sciences & Technology, Amgen Inc., Seattle, WA; and.
authors.forename: Heather
authors.initials: H
authors.lastname: Nunn
authors.affiliation: Small Molecule Process and Product Development, Amgen Inc., Thousand Oaks, CA.
authors.forename: Asher
authors.initials: A
authors.lastname: Lower
authors.affiliation: Product Attribute Sciences, Amgen Inc., Thousand Oaks, CA;
authors.forename: Gary
authors.initials: G
authors.lastname: Rogers
authors.affiliation: Drug Product Development, Amgen Inc., Thousand Oaks, CA; snashed@amgen.com.
authors.forename: Yasser
authors.initials: Y
authors.lastname: Nashed-Samuel
mesh_terms: D019149:Bioreactors; D004340:Drug Contamination / Q000517:prevention & control; D015195:Drug Design; D004345:Drug Industry; D004349:Drug Packaging; D009930:Organic Chemicals / Q000737:chemistry; D010969:Plastics / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D014867:Water / Q000737:chemistry
chemical_list: D009930:Organic Chemicals; D010969:Plastics; D011108:Polymers; D012997:Solvents; D014867:Water
keywords: Antioxidants; Extractables; Leachables; Plasticizers; Single-use bioreactor