Identification of a Leachable Compound Detrimental to Cell Growth in Single-Use Bioprocess Containers

Hammond, Michelle; Nunn, Heather; Rogers, Gary N.; Lee, Hans C.; Marghitoiu, Anatolia-Liliana; Pérez, Lourdes; Nashed-Samuel, Yasser; Anderson, Carl A.; Vandiver, Michael; Kline, Sally

doi:10.5731/pdajpst.2013.00905

Cited by 78 publications

(93 citation statements)

References 14 publications

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“…In shifting from polystyrene‐based flask cultures to bag systems, the profile of leachables and extractables is expected to change considerably due to the different types and quantities of plasticizers used in the manufacture of “plastic” disposable systems for cell culture. Leachables and extractables found in polystyrene or ethylene‐vinyl acetate polymers can have unknown, and both negative or positive effects on cell processing . Because extractables’ profiles may vary from lot to lot for some materials, extractables can reduce cell culture process reproducibility.…”

Section: Bioprocessing and Scale‐up Considerations In Bag Culture Sysmentioning

confidence: 99%

Bags versus flasks: a comparison of cell culture systems for the production of dendritic cell–based immunotherapies

et al. 2018

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In recent years, cell-based therapies targeting the immune system have emerged as promising strategies for cancer treatment. This review summarizes manufacturing challenges related to production of antigen presenting cells as a patient-tailored cancer therapy. Understanding cell-material interactions is essential because in vitro cell culture manipulations to obtain mature antigen-producing cells can significantly alter their in vivo performance. Traditional antigen-producing cell culture protocols often rely on cell adhesion to surface-treated hydrophilic polystyrene flasks. More recent commercial and investigational cancer immunotherapy products were manufactured using suspension cell culture in closed hydrophobic fluoropolymer bags. The shift to closed cell culture systems can decrease risks of contamination by individual operators, as well as facilitate scale-up and automation. Selecting closed cell culture bags over traditional open culture systems entails different handling procedures and processing controls, which can affect product quality. Changes in culture vessels also entail changes in vessel materials and geometry, which may alter the cell microenvironment and resulting cell fate decisions. Strategically designed culture systems will pave the way for the generation of more sophisticated and highly potent cell-based cancer vaccines. As an increasing number of cell-based therapies enter the clinic, the selection of appropriate cell culture vessels and materials becomes a critical consideration that can impact the therapeutic efficacy of the product, and hence clinical outcomes and patient quality of life.

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Section: Bioprocessing and Scale‐up Considerations In Bag Culture Sysmentioning

confidence: 99%

Bags versus flasks: a comparison of cell culture systems for the production of dendritic cell–based immunotherapies

et al. 2018

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“…The observation of reduced cell growth in media prepared from water held in certain bags clearly precludes adsorptive removal of key trace nutrients as a root cause and thus implicates the migration of one or more cytotoxic leachable compounds from the container and into the process liquid. Our company is one of those having instituted such a “cell‐based assay” (CBA) for plastic film process performance, and concurrently we have discovered and reported on the existence of a particular leachable compound that has cytotoxic effects when spiked into CHO cell culture even at relatively low concentrations . That compound, bis (2,4‐di‐ tert ‐butylphenyl) phosphate (bDtBPP, see Scheme ), is generated by the degradation of tris (2,4‐di‐ tert ‐butylphenyl)phosphite (TBPP, often referred to under the trade name Irgafos 168®), an antioxidant stabilizer that is commonly added to resin formulations of the kinds of polymers from which flexible bioprocess equipment is made.…”

Section: Introductionmentioning

confidence: 99%

A cytotoxic leachable compound from single‐use bioprocess equipment that causes poor cell growth performance

Hammond

Marghitoiu

Lee

et al. 2014

Biotechnology Progress

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A current trend in the production of biopharmaceuticals is the replacement of fixed stainless steel fluid-handling units with disposable plastic bags. Such single-use systems (SUS) offer numerous advantages, but also introduce a new set of materials into the production process and consequently expose biomanufacturers to a new set of risks related to those materials, not to mention reliance on an entirely new supply chain. In the course of developing and conducting a cell-growth-based test for suitability of disposable plastic components destined for use in cell culture operations, we discovered that the cytotoxic compound bis(2,4-di-tert-butylphenyl)phosphate (bDtBPP) leaches out of certain bags and into cell culture media in concentrations that are deleterious to cell growth. Specifically, media held in certain bags for several days at 37°C was found to contain bDtBPP, and use of those held-media samples in cell growth experiments provides data that overlap neatly with cell growth experiments using media spiked directly with bDtBPP, proving that bDtBPP leaching is responsible for the reduced growth attributable to those SUS bags. Overall, this issue represents a risk to the production of biopharmaceuticals in SUS, a risk that must be managed by diligent collaboration among companies along the entire supply chain for SUS components.

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“…Furthermore, in these complex mixtures, even constituents at low concentrations (low ppm to sub‐ppm) can potentially have pharmacological effects. Potential activity at low levels is why leachable constituents are an area of concern (Hammond et al, , ). The combination of chromatography with mass spectrometric detection provides the best analytical approach to investigate E&L, due to the inherent sensitivity and characterization capabilities.…”

Section: Applications Of Mass Spectrometrymentioning

confidence: 99%

The role of mass spectrometry and related techniques in the analysis of extractable and leachable chemicals

Sica

Krivos

Kiehl

et al. 2019

Mass Spectrometry Reviews

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In addition to degradation products, impurities, and exogenous contaminants, industries such as pharmaceutical, food, and others must concern themselves with leachables. These chemicals can derive from containers and closures or migrate from labels or secondary containers and packaging to make their way into products. Identification and quantification of extractables (potential leachables) and leachables, typically trace level analytes, is a regulatory expectation intended to ensure consumer safety and product fidelity. Mass spectrometry and related techniques have played a significant role in the analysis of extractables and leachables (E&L). This review provides an overview of how mass spectrometry is used for E&L studies, primarily in the context of the pharmaceutical industry. This review includes work flows, examples of how identification and quantification is done, and the importance of orthogonal data from several different detectors. E&L analyses are driven by the need for consumer safety. These studies are expected to expand in existing areas (e.g., food, textiles, toys, etc.) and into new, currently unregulated product areas. Thus, this topic is of interest to audiences beyond just the pharmaceutical and health care industries. Finally, the potential of universal detector approaches used in other areas is suggested as an opportunity to drive E&L research progress in this arguably understudied, under‐published realm.

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Identification of a Leachable Compound Detrimental to Cell Growth in Single-Use Bioprocess Containers

Cited by 78 publications

References 14 publications

Bags versus flasks: a comparison of cell culture systems for the production of dendritic cell–based immunotherapies

Bags versus flasks: a comparison of cell culture systems for the production of dendritic cell–based immunotherapies

A cytotoxic leachable compound from single‐use bioprocess equipment that causes poor cell growth performance

The role of mass spectrometry and related techniques in the analysis of extractable and leachable chemicals

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