Biomanufacturing using single use systems: Case study of fluoropolymer material

In this presentation, we will discuss recent biocompatibility data obtained using our gamma stable fluoropolymer platform and its advantage in a modern manufacturing environment for the handling of almost any biological fluids. Single-use, disposable solutions are now widely accepted as gold standard in the biopharmaceutical industry covering every single steps of the drug manufacture process. From early-stage small scale R&D project, upstream production with several thousand-liters bioreactors, downstream process, API formulation up to critical fill & finish and transport applications, the industry is taking a shift with a significant breakthrough reported in 2021: the largest GMP facility in the world using single-use (from 200L to 4,000 L) bioreactors has been publicly announced this year with a global single-use bioreactor workforce of over 150,000 L (with extension plans for 2024 to expand its use of disposable bioreactors to over 430,000 L). Please click Download on the upper right corner to see the full abstract

Adopting a fully single-use process to improve speed to clinic: A leachables case study

The implementation of single-use technologies for pharmaceutical product development continues to gain momentum; this trend is due to the advantages of increased flexibility, speed of implementation and lower capital investment In particular, they are seen as a means to accelerate the production of material for clinical trials. However, a primary concern regarding the use of such technologies is the impact leachables on patient safety in the final drug substance. Typically this concern is addressed through a patient safety evaluation utilizing extractable substances data based on model solvent extractions and from individual components and devices. However, little if any data has been published where leachables are evaluated under actual process conditions through a complete single-use clinical-scale process train. We have addressed this by completing a pilot scale 100 L “mock” mAb production and purification where the cells, and hence mAb protein, are absent but where the bioreactor was run for the normal duration with cell culture media and feeds, and the DSP train utilized all the standard process devices, buffers, conditions, and procedures. Data will be presented on the leachables profile throughout the entire production process the result of a patient safety evaluation conducted on the bulk drug substance after storage

Managing risk and consistency in the raw material supply chain for single use systems

As the bioprocessing industry moves increasingly towards single use systems (SUS), security of supply at the raw material level becomes imperative to quality. Long term supply of consistent, high quality materials desired for single use film is directly attributed to the stability, continuity and quality of raw material supply from the film manufacturer. This presentation will focus on the strategic selection of raw materials for a single use platform film in the bioprocessing industry to minimize risk associated with change notifications. Utilizing a quality by design approach, a single use film was formulated with no added Irgafos 168, enhanced abuse resistance and a low extractables and leachables profile. There are several factors that are critically important to understanding quality and risk of supply in raw material sourcing. This presentation will outline the raw material selection process established to ensure supply continuity and high quality desired for use in bioprocessing films. A comprehensive study was conducted on raw materials prior to film validation and quality controls were established to ensure consistency prior to processing into single use films

Analysis of leachable Bis Di-tert-butyl Phenyl Phosphate (bdtbpp) in bioprocessing films

The analysis of extractable and leachable compounds from medical grade plastics is a complex issue that is compounded by the presence of many chemical species that are either direct or indirect reaction and/or degradation products of additives, process aids and polymer agents. These compounds may be present at various levels and the potential for adverse effects on cell growth remains to be determined and warrants further investigation. Since many of these films undergo aggressive processing steps, such as, thermal extrusion and gamma irradiation there is the potential for many unknown degradation products to be formed at each step of the processing. In addition, the absolute identification of many of these chemical compounds remains unknown. Within the various applications of single-use disposable bioprocessing, the presence of bis di-tert butyl phenyl phosphate (bDtBPP), a common gamma irradiation degradation product of tris (2,4-ditert-butylphenyl) phosphite (TBPP), has been shown to have a profoundly negative impact on cell growth for certain lines. The presence of bDtBPP even at low levels (on the order of 10ppb) has been shown to inhibit cell growth performance percentages in some lines by as much as 30-50% [1]. The quantitative analysis of this compound becomes increasingly difficult at lower levels due to either; 1) irreversible binding of strongly charged phosphate groups to glassware and other labware used in processing samples, or 2) hydrolytic degradation in aqueous solutions, or 3) any combination of the two. Losses from either of these conditions has been show to give rise to variations in quantitative analysis results as high as 50% when testing ranges are set between 5-100 parts per billion (ppb) in water. To this end, we have investigated the fate of this compound at part per billion levels to gain insight into possible mechanisms associated the variations observed. The hydrolytic degradation as well as irreversible binding to various substrates such as HPLC vials, pipette tips, etc. has been investigated extensively. We propose mitigation strategies which allow for low level quantitative analysis of this compound to achieve a coefficient of variation (CV) within the range of 10-20%, for bDtBPP in water within concentration ranges of 5-100 ppb. References Lindskog, Eva., Blank, Eva., Ullsten, Sara., Yi, Shujian., Ganguli, Pokon., Carter, Jeffrey., Parma, Hernan. Implementation of Raw Material Control Strategies in the Manufacture of Single-Use Bioprocessing Containers; BioPharm International, Volume 28, Issue 1, January 1, 2015

Flexural fatigue behavior of rocking bioreactor films

The fields of biopharmaceutical processing and cell therapy are adopting single-use, closed systems throughout their workflows to enhance sterility, minimize waste wash effluent and enable manufacturing flexibility compared to traditional stainless steel bioreactors. One of the key single-use technologies in use is the rocking bioreactor, comprising a polymer film bag (outfitted with ports and sensors) mounted to a tray capable of mixing the contents of the bag and a control system (controlling temperature, agitation, and potentially media perfusion). One of the challenges encountered in rocking bioreactor bags is the fact that upon inflation/filling with media, the originally flat bioreactor bags often develop folds and dimples due to their inflated geometry. These deformations tend to be inconsequential at small volumes and low agitation rates/times, but can lead to flex fatigue failures such as whitening, delamination and through-cracking under more extreme conditions. In practice, these failures are dependent on a number of factors including bag material and volume, mounting geometry, rocking angle and rate, and the duration of culture, making a systematic study of the material properties controlling this behavior difficult and time-consuming. Several flex fatigue testing systems exist in the literature, including Gelbo and Sonntag-Universal, but none of these effectively model the unique geometry and stresses of the rocking bioreactor geometry. To this end, we have developed accelerated test methods to analyze the flexural fatigue behavior of multilayer rocking bioreactor films. These methods enable quality control testing of film lots, and have the potential to compare different film compositions with a rapid and reproducible test, thereby facilitating development of new films. Our test method models the local geometry surrounding the fold/dimple in a rocking bioreactor in a small sample of film, and cycles the sample to accelerate flexural fatigue at the dimple site. Initial results indicate the ability to accelerate film failure from tens of days on a rocking bioreactor platform (using a full bioreactor bag) to tens of hours using less than ten square inches of film. We will discuss the effects of various experimental parameters on film failure, optimization of test procedures and correlation with rocking bioreactor testing in the field

Test method development for next-generation bioprocessing applications

As single use disposable (SUD) bioprocessing systems become more commonplace, the range of applications and workflow steps served by single use continues to grow. Nearly all of the process steps and techniques currently used in bioprocessing were developed on stainless steel or glass vessels which exhibit thermal, chemical and mechanical properties which differ greatly from the polymer films used in SUD. While in many cases (e.g., cell culture) these differences have negligible effects on performance, as the industry pushes the limits of intensification and increases in the breadth of SUD applications (antibody-drug conjugation – ADC, microbial culture, etc.) single-use materials and systems face new challenges. This highlights the need for testing capabilities to qualify and develop new SUD materials, components and systems to ensure integrity, cleanliness and performance. In this presentation, we will comment on the current state-of-the-art in standardized testing as well as highlighting several small-scale test methods developed internally, specifically for testing materials for SUD bioprocessing. In order to assess film and component (e.g., port, tubing) capabilities for the full range of current and foreseeable process steps, we have developed/adapted/adopted test methodologies to model environments experienced both in conventional use (abrasion, flexure, pressurization), as well as challenging new use cases including high and low temperature extremes (freezing, pasteurization) and aggressive, non-aqueous environments used in bioconjugate chemistry (e.g., ADC, conjugate vaccines), chemical viral inactivation and non-traditional cell culture (e.g., microbial). For each of these applications, our methods were developed based on generating an understanding of the environment in which the chosen SUD system is to be used, identifying the primary failure modes that are (or could be) encountered in use and reducing these risks to their fundamental physics to generate small-scale tests that can be performed rapidly on small samples of material. These test methods allow for rapid screening of film and component materials to reduce risks in new applications prior to prototype development and assess product and process quality in the long ter