Single-use Technology Research Articles

Multi-stage and multi-objective decision-support tool for biopharmaceutical drug product manufacturing: Equipment technology evaluation

The emergence of single use technology (SUT) for equipment promises to be a revolutionary change in biopharmaceutical production. SUT equipment reduces non-value adding working time and provides operational flexibility. However, there are increased process risks such as possible contamination with leachable compounds or supply dependencies, as well as required redesign efforts. This work provides a multi-stage approach for decision-support for optimal equipment technology applied in a case study for a sterile filling line. Three design stages are identified. The first two stages explore the design landscape using the economic performance to show the general trends of favorable regions for equipment technology and the sensitivity to process parameters according to their relative importance. The last stage is more detailed but incorporates additional assessment indicators such as environmental impact and risks of process downtime. From an economic perspective SUT options are more favorable for short term and small-scale production, such as the case for preparing drugs for clinical trials. Traditional stainless-steel options had a significant economic advantage at large scale and long term production. The ranking of favorable options varied as other assessment factors, namely life-cycle CO2 emissions as well as risks in contamination and supply delay, were included in the analysis. The proposed methodology offers an agile decision-making tool, accounts for uncertainties across various design phases, and offers a systematic exploration of the available design landscape.

Identification of additives in polymers from single-use bioprocessing bags by accelerated solvent extraction and ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry

Single-use technologies are increasingly used in biopharmaceutical manufacturing. Despite their advantages, these plastic assemblies draw concern because they are a potential source of contamination due to extractable and leachable compounds (E&Ls). Characterising E&Ls from such materials is a necessary step in establishing their suitability for use. Therefore, there is an urgent need for sensitive methods to identify and quantitatively assess compounds in plastic materials.Accelerated solvent extraction (ASE) is a powerful technique that can be reliably used for this purpose. In this study, ASE followed by liquid chromatography and Orbitrap-based High Resolution Accurate Mass (HRAM) mass analysis was found to be an efficient and versatile method for the determination of additives in different multilayer polymer systems from single-use bags. ASE optimisation was performed using a design of experiments approach. The type of solvent, temperature, swelling agent addition, static time and number of cycles were the selected variables. Optimum conditions were dependent on the type of plastic film. Ethyl acetate and cyclohexane were selected individually as optimum solvents. Optimum temperatures were 90–100 °C. Pressure was set at 1500 psi and extraction time was 30 min in 2 cycles. Swelling agent addition was necessary with polar extraction solvents. More than 100 additives and degradation products were confidently identified by HRAM MS. Correlations between the type and levels of identified additives and the type of polymer system were established. In addition, degradation behaviour and pathways for some additives can be addressed.

Maximizing speed to market: flexibility benefits of single-use technology for gene therapy manufacture

Maximizing speed to market: flexibility benefits of single-use technology for gene therapy manufacture

Estimating capital investment and facility footprint in cell therapy facilities

Estimations of the facility footprint and fixed capital investment (FCI) of cell therapy (CT) facilities need to consider the unique features of the single-use technologies (SUTs) selected for CT manufacture (e.g. cleanroom containment requirement, capacity, automation) and the product nature that impacts scale-out versus scale-up approaches. A novel detailed factorial methodology is proposed for estimating FCI and footprint for bespoke stick-built cell therapy facilities that accounts for technology-specific factors for key cell culture technologies as well as the implications of SUTs, open versus closed operations and the commercialisation scenario selected. This was used to derive benchmark values for short-cut cost and area factors for typical cell therapy facilities according to the technologies selected. The results provide project-specific ratios for equipment purchase costs to facility footprint (area factor) and for FCI to total equipment purchase costs (cost factor or “Lang” factor). Area factors ($/m2) were 675-6,815 and the cost factors were 2.3–8.5 for a greenfield project in a medium-developed country. The case study shows that for the same output, facility footprints and FCI values are on average 6 times higher for autologous processes than allogeneic processes. This is attributed to economies of scale achieved with scale-up for allogeneic cell therapy manufacture. This study can be used to predict the commercial FCI and facility footprint during early stages of process development.

Single-use, single-pass tangential flow filtration using low-cost hollow fiber modules

Increasing pressures on biomanufacturing costs have led to renewed interest in the development of single-use technologies that can be readily adapted to continuous processing. The objective of this study was to use commercially available hollow fiber membranes, originally designed for high-flux hemodialysis, for single pass tangential flow filtration with high conversions. Experiments were performed with solutions of Immunoglobulin G (IgG) using polysulfone hollow fiber membrane cartridges with a surface area of 1.9 m2. The hollow fiber modules were able to provide more than 10-fold concentration of IgG in a single-pass with exit concentrations greater than 200 g/L. Stable operation was achieved during a continuous run for over a 120-h period with feed side pressure drops less than 1 kPa and transmembrane pressure drops less than 60 kPa. A single dialyzer was able to process 1.4 kg of IgG per day with a total cost of less than $0.004 per g of IgG. Flux-pressure profiles were in good qualitative agreement with a simple model accounting for concentration polarization effects and the variation of flux and pressure with axial position in the module. These results clearly demonstrate the potential of using these low-cost single-use hollow fiber modules for single-pass tangential flow filtration in continuous biomanufacturing.

Online Decision-Support Tool “TECHoice” for the Equipment Technology Choice in Sterile Filling Processes of Biopharmaceuticals

In biopharmaceutical manufacturing, a new single-use technology using disposable equipment is available for reducing the work of change-over operations compared to conventional multi-use technology that use stainless steel equipment. The choice of equipment technologies has been researched and evaluation models have been developed, however, software that can extend model exposure to reach industrial users is yet to be developed. In this work, we develop and demonstrate a prototype of an online decision-support tool for the multi-objective evaluation of equipment technologies in sterile filling of biopharmaceutical manufacturing processes. Multi-objective evaluation models of equipment technologies and equipment technology alternative generation algorithms are implemented in the tool to support users in choosing their preferred technology according to their input of specific production scenarios. The use of the tool for analysis and decision-support was demonstrated using four production scenarios in drug product manufacturing. The online feature of the tool allows users easy access within academic and industrial settings to explore different production scenarios especially at early design phases. The tool allows users to investigate the certainty of the decision by providing a sensitivity analysis function. Further enrichment of the functionalities and enhancement of the user interface could be implemented in future developments.

Biopharma Keeps Shifting the Steel–Plastic Balance

Genetic Engineering & Biotechnology NewsVol. 39, No. S3 Biopharma Keeps Shifting the Steel–Plastic BalanceBiopharma is stepping up its adoption of single-use technology—not just in preclinical and clinical development, but in commercial operationsGareth John MacdonaldGareth John MacdonaldSearch for more papers by this authorPublished Online:29 May 2019https://doi.org/10.1089/gen.39.S3.02AboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View articleFiguresReferencesRelatedDetails Volume 39Issue S3Jun 2019 InformationCopyright © by GEN PublishingTo cite this article:Gareth John Macdonald.Biopharma Keeps Shifting the Steel–Plastic Balance.Genetic Engineering & Biotechnology News.Jun 2019.S4-S7.http://doi.org/10.1089/gen.39.S3.02Published in Volume: 39 Issue S3: May 29, 2019PDF download

Improvements in single‐use bioreactor film material composition leads to robust and reliable Chinese hamster ovary cell performance

Single-use technologies, in particular disposable bioreactor bags, have become integral within the biopharmaceutical community. However, safety concerns arose upon the identification of toxic leachable compounds derived from the plastic materials. Although the leachable bis(2,4-di-tert-butylphenyl)-phosphate (bDtBPP) has been previously shown to inhibit CHO cell growth, it is critical to determine if other compounds like this are still present in subsequent generations of films for industrial application. This study compares the performance of CHO cells, CHO-K1, and CHO-DP12, cultured in media conditioned in an older single-use bioreactor (SUB) film (F-1) and a newer generation film (F-2) from the same vendor. CHO cells cultured in media conditioned for 7 days in the F-1 film demonstrated significantly reduced growth and antibody productivity profiles when compared to controls and media conditioned for the same time period in the newer F-2 film. Proteomic profiling of CHO cells cultured in the F-1 conditioned media identified differentially expressed proteins involved in oxidative stress response as well as compromised ATP synthesis. These potentially metabolically compromised cells exhibited reduced oxidative phosphorylation activity as well as lower glycolytic metabolism, characteristic of slower growing cells. Nonvolatile and metal leachables analysis of film extracts by LC-MS revealed a reduction in the abundance of the analyzed leachates from F-2 films when compared to F-1 films including bDtBPP, potentially explaining improved CHO cell growth in F-2 conditioned media. Furthermore, in vitro endocrine disruptor testing of the known leachable revealed this molecule to possess the potential to act as an androgen antagonist. This study demonstrates an improvement in the materials composition used in modern generations of SUBs for safe application in the bioprocess.

Impact of depth filtration on disulfide bond reduction during downstream processing of monoclonal antibodies from CHO cell cultures.

Monoclonal antibody interchain disulfide bond reduction was observed in a Chinese Hamster Ovary manufacturing process that used single-use technologies. A similar reduction has been reported for processes that involved high mechanical shear recovery unit operations, such as continuous flow centrifugation and when the clarified harvest was stored under low dissolved oxygen (DO) conditions (Trexler-Schmidt et al., 2010. Biotechnology and Bioengineering, 106(3), 452-461). The work described here identifies disposable depth filtration used during cell culture harvest operations as a shear-inducing unit operation causing cell lysis. As a result, reduction of antibody interchain disulfide bonds was observed through the same mechanisms described for continuous flow centrifugation. Small-scale depth-filtration models were developed, and the differential pressure (Δ P) of the primary depth filter was identified as the key factor contributing to cell lysis. Strong correlations of Δ P and cell lysis were generated by measuring the levels of lactate dehydrogenase and thiol in the filtered harvest material. A simple risk mitigation strategy was implemented during manufacturing by providing an air overlay to the headspace of a single-use storage bag to maintain sufficient DO in the clarified harvest. In addition, enzymatic characterization studies determined that thioredoxin reductase and glucose-6-phosphate dehydrogenase are critical enzymes involved in antibody reduction in a nicotinamide adenine dinucleotide phosphate (NADP + )/NADPH-dependent manner.

Emerging Trends in Single-Use Technology in the Manufacturing of Antibody-Drug Conjugates

Single-use technology, designed for the manufacturing of biopharmaceutical products, has made major inroads over the last 30 years. First introduced in the late 1970s in the form of disposable capsules and a range of filters, single-use technologies were revolutionized in the late 2000s with the introduction of single-use 2D and 3D process containers and filter assemblies for mixing and storage systems. Today, these technologies have been adopted across the upstream manufacturing process, downstream purification and fill-finish of entire classes of biologic drugs.

Large-Scale Assessment of Extractables and Leachables in Single-Use Bags for Biomanufacturing.

Single-use technologies (SUTs) are widely used during biopharmaceutical manufacture as disposable bioreactors or media and buffer storage bags. Despite their advantages, the risk of release of extractable and leachable (E&Ls) substances is considered an important drawback in adopting disposables in the biomanufacturing process. E&Ls may detrimentally affect cell viability or productivity or may persist during purification and present a risk to the patient if remaining in the final drug product. In this study, 34 plastic films from single-use bags (SUBs) for cell cultivation were extracted with selected solvents that represent reasonable worst-case conditions for most typical biomanufacturing applications. SUBs were incubated at small-scale under accelerated-aging conditions that represented standard operational conditions of use. Leachables analysis was performed following dispersive liquid-liquid microextraction (DLLME) for analyte preconcentration and removal of matrix interference. Resulting extracts were characterized by GC-headspace for volatiles, high resolution GC-Orbitrap-MS/MS for semivolatiles, high resolution LC-Orbitrap-MS/MS for nonvolatiles, and ICP-MS for trace elemental analysis. Multivariate statistical analysis of the analytical data revealed significant correlations between the type and concentration of compounds and bags features including brand, manufacturing date and polymer type. The analytical data demonstrates that, over recent years, the nature of E&Ls has been altered due to the implementation of manufacturing changes and new types of polymers and may change further with the future advent of regulations that will limit or ban the use of certain raw materials and additives. The broad E&L database generated herein facilitates toxicological assessments from a biomanufacturing standpoint and provides practical guidelines for confident determination of E&Ls to enable screening and elimination of nonsatisfactory films for single use bioprocessing.

Multiobjective decision-support tools for the choice between single-use and multi-use technologies in sterile filling of biopharmaceuticals

In sterile filling of biopharmaceuticals, two equipment technologies are available, namely, a conventional multi-use technology using stainless steel fixed facilities, and a new single-use technology using resin-made disposable equipment. For the choice between these technologies, this study proposes a set of three multiobjective decision support tools. The first tool is to evaluate cost, environmental impact, product quality, and supply robustness; the second uses a set of weighting factors to produce a total score; the third conducts a sensitivity analysis to investigate the influence of the weighting factors on the final decision. The use of these tools was described as an activity model by a method called "the type zero method of integration definition for function modeling" (IDEF0). A case study was conducted to demonstrate the tools and the activity model in different production patterns, i.e., from small-scale and multiproduct to large-scale and single-product.

Computational studies of a novel magnetically driven single-use-technology bioreactor: A comparison of mass transfer models

This work applies computational fluid dynamics (CFD) modelling to a novel 1000 L design of single-use-technology (SUT) bioreactor, with a magnetically driven floor-mounted impeller and spargers distributed across the tank floor. A two-phase Euler-Euler model using the k-ε turbulence model and population balance is presented alongside the use of immersed solid method for modelling the impeller motion. This work also provides the first CFD analysis of a large-scale SUT bioreactor, identifying key flow characteristics of the non-standard design at different operating conditions. Five models for the mass transfer coefficient, kL, are compared, with kLa values compared to experimental measurements. The slip velocity model is found to be the best prediction of the mass transfer coefficient for this SUT system. Separating the influence of the mass transfer coefficient and specific area,a shows that the latter is the dominant driving force behind changes in kLa that occur at different operating conditions. Comparing the present work to previous studies for traditional stirred tanks highlights the need for understanding the hydrodynamics of non-standard reactor designs when identifying suitable mass transfer models in gas–liquid flow systems.

DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein.

The transmembrane rabies virus glycoprotein (RVGP) is the main antigen of vaccine formulations used around the world to prevent rabies, the most lethal preventable infectious disease known. The objective of this work was to evaluate the potential of a bioreactor using wave-induced agitation in the initial steps of scaling up the rRVGP production process by a Drosophila melanogaster S2 cell line to produce rRVGP in sufficient quantities for immunization and characterization studies. Taking advantage of some remarkable features recognized in Drosophila S2 cells for scaling the culture process, a robust recombinant lineage (S2MtRVGPH-His) engineered by our group for the expression of rRVGP using a copper-inducible promoter was used in the bioreactor cultures. The WAVE Bioreactor was chosen because it represents an innovative approach to the cultivation of animal cells using single-use technology. For that purpose, we firstly established a procedure for culturing the S2MtRVGPH-His lineage in 100 mL Schott flasks. Using an inoculum of 5 × 105 cells/mL in culture medium (Sf900-III) induced with solution of CuSO4 (0.7mM) and a convenient pH range (6.2-7.0), optimal parameter values such as time of induction (72h) and temperature (28°C) to increase rRVGP production could be defined. This procedure was reproduced in culture experiments conducted in a WAVE Bioreactor™ 2/10 using a 2 L Cellbag. The results in Schott flasks and in WAVE Bioreactor™ were very similar, yielding a maximum titer of rRVGP above of 1 mg.L-1. The immunization study showed that the rRVGP produced in the bioreactor was of high immunogenic quality.

Development of a 3-step straight-through purification strategy combining membrane adsorbers and resins.

The pressures to efficiently produce complex biopharmaceuticals at reduced costs are driving the development of novel techniques, such as in downstream processing with straight-through processing (STP). This method involves directly and sequentially purifying a particular target with minimal holding steps. This work developed and compared six different 3-step STP strategies, combining membrane adsorbers, monoliths, and resins, to purify a large, complex, and labile glycoprotein from Chinese hamster ovary cell culture supernatant. The best performing pathway was cation exchange chromatography to hydrophobic interaction chromatography to affinity chromatography with an overall product recovery of up to 88% across the process and significant clearance of DNA and protein impurities. This work establishes a platform and considerations for the development of STP of biopharmaceutical products and highlights its suitability for integration with single-use technologies and continuous production methods. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:931-940, 2017.

Erratum to: Decision-Support Method for the Choice Between Single-Use and Multi-Use Technologies in Sterile Drug Product Manufacturing

Single-use technology has been applied to sterile drug product manufacturing processes as a new technology in contrast with the conventional multi-use technology. This study proposes a decision-support method for choosing between these two technologies in sterile drug product manufacturing. The proposed method consists of four steps: create process evaluation models, calculate evaluation results, perform what-if analysis, and interpret results. We created models for evaluating the production cost and life cycle CO2 emissions of processes using either technology. “What-if” analysis quantifies the effect of the input parameters on the evaluation results, which supports more informed decision-making. Here, it is recommended that the filling speed, which was found to have a significant impact on filling accuracy—a critical quality attribute of sterile drug products—should be investigated. As a case study, the method was applied in two cases of technology selection: (i) single-batch production of a product with different batch sizes and (ii) single-batch production with different production patterns. The single-use technology showed its economic superiority in producing small batches and in producing multiple small-scale products, whereas in the environmental evaluation, it was always better than multi-use technology. What-if analyses revealed the impact of changing input parameters on the economy, environment, and quality. Our method can support the choice of single-use and multi-use technologies in plants having both technologies independently. In the case study, economic evaluation showed a critical point in each design case, whereas the environmental evaluation result was always better in single-use technology.

Non-volatile extractable analysis of prefilled syringes for parenteral administration of drug products

The determination of extractable profiles for single-use technologies represents an important aspect of pharmaceutical production to minimize any possible compromise in drug product quality or potential risk to patients by identifying substances that may potentially leach from such devices. An approach for the extractable assessment of prefilled syringes, a promising alternative for parenteral administration of pharmaceutical products, is described herein. Four extraction solvents were selected: a mixture 2-propanol:water (1:1), was intended to represent aggressive conditions to extract a broad spectrum of extractables, including organic additives and substances which are poorly water-soluble. Extractions with buffers at three different working pH values spanning a range standardly used in pharmaceutical formulations were also evaluated to identify substances that require specific conditions for their extraction due to their individual chemical properties. Syringes from two different brands were analysed along with their corresponding plunger stoppers. Syringes were extracted at 40°C for 4days, the plunger stoppers were extracted with 2-propanol at 70°C for 24h according to ISO 10993-12:2012. Extractables were identified by UHPLC–MS on a quadrupole time of flight instrument using a non-targeted discovery strategy. A total of 25 compounds were identified, mostly polymer additives and their degradation products. The presented methodology represents a reference point for further studies focused on the characterisation of extractables and leachables from prefilled syringes.

Thermo Fisher buys process systems maker

Thermo Fisher Scientific plans to buy Finesse Solutions, a maker of process sensors, controllers, and software for biopharmaceutical production with annual sales of $50 million. Terms of the transaction were not disclosed. A key motivator behind the acquisition is Finesse’s development of a universal control system that “will combine seamlessly with our existing single-use technologies,” Thermo Fisher CEO Marc N. Casper says.

Early Assessment of the Likely Cost Effectiveness of Single-Use Flexible Video Bronchoscopes.

BackgroundBronchoscopic procedures are common in the clinical setting, with estimates indicating 500,000 are undertaken per year in the USA alone. These procedures are generally regarded as safe. Unfortunately, a risk of cross-contamination between patients, with possible subsequent infection, is associated with the re-usable technology typically used in these procedures.ObjectiveOur objective was to conduct an early cost-effectiveness analysis (CEA) of single-use flexible video bronchoscope technology compared with the current reusable technology in a US hospital intensive care setting.MethodsWe conducted a CEA to determine an incremental cost-effectiveness ratio (ICER), and constructed a decision analytic model based on the best available evidence from a literature search and a Delphi panel. We also conducted several one- and two-way sensitivity analyses and a probabilistic sensitivity analysis to illuminate the uncertainty associated with the estimates.ResultsThe literature search showed ample evidence of risk, albeit little of it was quantifiable. Estimates from the Delphi method found approximately a 3% risk of cross-contamination and approximately a 21% risk of subsequent infection. Pneumonia was estimated as the most likely manifestation of infection. The CEA showed a saving of $US118 per procedure and elimination of 0.7% of the risk of infection with the single-use technology. Relevant sensitivity analyses generally validated this result.ConclusionThis study suggests that implementation of the single-use technology in the intensive care unit is cost effective in most scenarios. However, this result should be interpreted with caution because of the lack of certain knowledge on this particular topic.Electronic supplementary materialThe online version of this article (doi:10.1007/s41669-017-0012-9) contains supplementary material, which is available to authorized users.

Fully Disposable Manufacturing Concepts for Clinical and Commercial Manufacturing and Ballroom Concepts.

The availability and use of pre-sterilized disposables has greatly changed the methods used in biopharmaceuticals development and production, particularly from mammalian cell culture. Nowadays, almost all process steps from cell expansion, fermentation, cell removal, and purification to formulation and storage of drug substances can be carried out in disposables, although there are still limitations with single-use technologies, particularly in the areas of pretesting and quality control of disposables, bag and connections standardization and qualification, extractables and leachables (E/L) validation, and dependency on individual vendors. The current status of single-use technologies is summarized for all process unit operations using a standard mAb process as an example. In addition, current pros and cons of using disposables are addressed in a comparative way, including quality control and E/L validation.The continuing progress in developing single-use technologies has an important impact on manufacturing facilities, resulting in much faster, less expensive and simpler plant design, start-up, and operation, because cell culture process steps are no longer performed in hard-piped unit operations. This leads to simpler operations in a lab-like environment. Overall it enriches the current landscape of available facilities from standard hard-piped to hard-piped/disposables hybrid to completely single-use-based production plants using the current segregation and containment concept. At the top, disposables in combination with completely and functionally closed systems facilitate a new, revolutionary design of ballroom facilities without or with much less segregation, which enables us to perform good manufacturing practice manufacturing of different products simultaneously in unclassified but controlled areas.Finally, single-use processing in lab-like shell facilities is a big enabler of transferring and establishing production in emergent countries, and this is described in more detail in 7. Graphical Abstract.