Engineering Commons^™

Challenges And Status: Single-Use Bioreactors For Microbial Processes, Peter Neubauer, Anna Maria Marba Ardebol, Friederike Hillig, Olivia Zakrzewski, Jorn Emmerich, N.M.G. Ooosterhuis, S. Junne

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Nowadays single-use bioreactors are fully accepted in the biopharmaceutical industry. Reactors up to 2000L working volume are commonly used. However, these bioreactors are limited in terms of mass-transfer and mixing capabilities and therefore only suited for application in mammalian cell culture. Single-use processing offers many benefits like cost reduction, flexibility, reduction of contamination risks, etc. These benefits apply for both microbial processes as for mammalian processes.

Additionally, for the use of marine microorganisms the application of single-use bioreactors (SUB) offers a possibility to circumvent problem of corrosion which occurs in steel bioreactors due to the high chloride ion concentration in …

Applying Practices From The Medical Device Industry To Ensure Patient Safety Of Single-Use Products In The Biotechnology Industry, Jan Oberdoerster

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Biocompatibility (i.e., the quality of not having toxic or injurious effects on biological systems) requirements for single-use products in the biotechnology industry can vary depending on point of use. For example, single-use syringes are considered a combination medical device and thus require assessment according to the ISO 10993 standards with test ranging from acute systemic toxicity to genotoxicity to sensitization. Processing equipment (e.g., reaction vessels, tubing, filters, chromatography columns) in contrast, generally requires a USP Class VI designation (a battery of in vivo tests consisting of acute systemic toxicity, intracutaneous reactivity, and muscle implantation) with rigorous requirements for leachates. The …

Homogenizing Biologic Drug Substance Bulk Using Single Use Mixing Systems, Benson Gikanga, Yufei Chen, Oliver Stauch, Yuh-Fun Maa

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Mixing of biologic Drug Substance (DS) is a critical process step during manufacturing of biologic Drug Product (DP). Generally, mixers used are equipped with either bottom-mounted or top-mounted agitators. Bottom-mounted mixers (BMMs), particularly those that are magnetically driven, are preferred because of their associated low risk of contamination, ease of use, and ability to accommodate low minimum mixing volumes. Despite these benefits the mechanical coupling of the rotating impeller and the male bearing on the tank may lead to shearing of monoclonal antibody (mAb) formulations generating elevated amount of particles in the DS or DP solution. This may lead to …

Successfully Employing Single-Use Bioreactors For Different Expression Systems, Valentin Jossen, Regine Eibl, Dieter Eibl

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

The single-use bioreactor market was estimated to have grown at a CAGR of 20% in 2014. Wave-mixed bioreactors and stirred bioreactors represent the largest segment of today`s single-use bioreactor market, and are preferred by process developers and manufacturers of preclinical and clinical samples when high-value products up to medium scale and products requiring high safety demands are in focus. However, wave-mixed and stirred single-use bioreactors are also becoming increasingly important for commercial biopharmaceutical productions. This is ascribed to the availability of high-productivity cell lines, which require smaller bioreactors. In addition, the availability of bioengineering data and cultivation bags with improved …

Single-Use Bioreactor Technologies For Early Stage Development Of Microalgae Cultivation Processes, Gary Lye, Ebenezer Ojo, Hadiza Auta, Frank Baganz

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

This work describes the engineering characterization and evaluation of two novel, single-use photobioreactor technologies for microalgal cultivation. There is currently considerable interest in microalgae as alternative expression systems for the production of value-added chemicals as well as therapeutic proteins and vaccines. In contrast to mammalian expression systems, however, bioreactor platforms to address early stage development challenges are not well established particularly for phototrophic and mixotrophic cultivation strategies.

To support early stage cell line selection and process characterization a single-use, 24-well micro photo-bioreactor (mPBr) was established together with an illuminated and environmentally controlled shaker platform [1]. The same orbital shaker platform …

Single Use Polymeric Materials: The Science Of Safety, Mitchell Cheeseman

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

No abstract provided.

Cell Culture Scale Translation From A 24-Well Single-Use Miniature Bioreactor And Subsequent Impact On Product And Broth Quality, John Betts, Franc Baganz, Gary Lye, Steve Warr, Gary Finka, Mark Uden

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

To accelerate cell culture process development, most companies have validated scale-down models of their pilot and manufacturing scale bioreactors. Advancing such mimics to even smaller scales requires the large scale engineering environment to be accurately recreated. Here we describe a single-use microwell methodology that accurately reproduces not only cell growth kinetics but also key attributes related to product quality and broth processability.

The μ24 miniature bioreactor system enables system level control of agitation (by orbital shaking), with individual well control of pH, DO and temperature. Two distinct plate types are investigated, allowing for either headspace or direct gas sparging. An …

Robust, Fast And Accurate Ph Sensing Platform Solution Targeting Single Use Application, Srinivas Rao, Hardjinder Ladhar, Jai Krishnamurthy

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

QbD in biotech and life-sciences necessitates good process metrics in drug discovery, development and manufacturing. Accurate knowledge of pH measurements remains fundamentally important. There is much to be desired with use of conventional glass potentiometric probes for monitoring pH. Challenges being measurement accuracy, system drift, and need for frequent calibration. Additionally glass being fragile compounds the situation further in manufacturing.

Glass probes for pH measurement are really unattractive for single use system applications (SUS). Optical based solutions while easy to integrate lack range, need calibration and are a problem in storage (application timing).

We demonstrate a robust Platform Solution tested …

Grey Space Bioprocessing: Myth Or Reality?, Diana Lok, Ekta Mahajan

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

As disposable skids are more frequently used in biotech companies and their comfort increases, their applications have slowly surpassed research and development into clinical and commercial products that impact human patients.

As the biotech market continues to grow, there is interest in grey space processing due to lower initial capital and operating costs. The aspiration is to be able to perform processing in grey space to reduce facility requirements while maintaining product purity. This poster will address

1) What defines grey space

2) How single use technology can facilitate closed processing

3) Can we leverage single use closed processing to …

Risk Reduction In Single-Use Assemblies By Implementing Helium Integrity Test And A New Point-Of-Use Leak Test Using Palltronic® Flowstar Lgr Instrument For Critical Formulation And Filling Applications, Anil Kumar, Vishwas Pethe, Ralf Kuriyel, Helene Pora

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

This presentation will focus on challenges with implementation of single use, including robustness concerns, ensuring closures, integrity of multi-polymer systems both during manufacturing as well as at point of use. Despite numerous challenges, rapid adoption of single-use technologies still continues due its application in both more critical applications and expansion to commercial scale. More particularly, single use bioprocessing assemblies (Biocontainers and multi component single use systems) are being utilized in increasingly critical and demanding applications such as for holding and transporting sterile bulk drug substance and formulation and final filling. Therefore, potential of product loss and operator safety risk, have …

Single-Use Bioprocess Containers: Raw Material Control Strategy For Optimal Cell Growth, Susan Burke, Sara Ullsten, Eva Blanck, Eva Lindskog, Shujian Yi, Jeffrey Carter, Pokon Ganguli, Herman Parma

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

There is an upward trend in the use of disposable containers for manufacturing biopharmaceuticals. Single-use systems offer several advantages such as increased flexibility, the elimination of cleaning procedures, and reduced capital expenditure. However, due to the plastic nature of these disposable bags, the materials of construction can be more complex than traditional stainless steel vessels. They are often made of films containing multiple layers of polymers with various additives for processing. There have been scientific reports of the inhibition of biopharmaceutical cell culture growth attributed to a leachate originating from an antioxidant commonly used in the plastic films of single-use …

Evaluation Of Single-Use Bioreactors For The Production Of A Hepatitis C Vaccine Candidate, Ricardo Silva, Joao Clemente, Rute Castro, Lenaig Savary, Alex Xenopoulos, Cristina Peixoto, Manuel Carrondo, Paula Alves

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

During the last decade, the usage of single-use bioreactors has been increasing in the biopharmaceutical industry. This technology offers some appealing advantages over their conventional counterparts made of glass or stainless steel, such as operational flexibility, faster batch-to-batch turnaround times and the reduction of clean-up and validation characteristic of single-use materials.

The present work examines a stirred single-use bioreactor for its suitability for the production of a Hepatitis C Virus-Like Particles (VLPs) vaccine candidate using the baculovirus expression system with Sf9 cells.

In this sense a 2L glass stirred tank and a Mobius^® 3L Bioreactor are compared in terms …

Identification Of Antioxidant By-Products Based On Their Specific Chemistry And Their Potential Detection During Sus Extractable Study, Samuel Dorey, Fanny Gaston, Nathalie Dupuy, Sylvain Marque, Gerard Audran

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Single-use films in biopharmaceutical and biotechnology industries are mainly made from polymers such as PE, EVA and EVOH. Depending upon the environmental aggressiveness during various stages of the polymer lifetime, additives are added to protect them such as substituted hindered phenols acting as antioxidants, melt (processing) stabilizers, and to some extent as photo-antioxidants.

Ionizing radiation effects on polymers have been widely investigated. They consist mainly of free radicals production. These free radicals can in turn lead to degradation and or crosslinking phenomena (release of gases, discoloration, changes in mechanical properties and gas permeability, degradation and leaching of polymer additives into …

Single Use Bioreactors: Geometry Does Matter, Colin Jaques, Anthony Beaney

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

The first generation of single use bioreactors (SUBs) departed from conventional stirred tank bioreactor (STR) geometry in terms of impeller number, and orientation and sparger hole diameter. Moreover, one marked feature of SUB bioreactors was that they could be operated at lower volumes than conventional STRs, bringing considerable operational flexibility. This practice, however, further negates the principle of geometric similarity. Whilst some processes may be able to remain within a SUB for the whole product life cycle, many products will require scale up to larger-scale vessels as demand for the product increases. This poster considers the implications of changing reactor …

Implementation And Characterization Of Solvent Detergent Viral Inactivation In Single Use Bags, Michael Cunningham, Ven Raman, Nitin Sood, Jessica Shea, Lori Mullin, Eric Youssef, Pedro Almeida, Elizabeth Goodrich

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Biomanufacturing of recombinant products from mammalian cell culture systems and plasma derived products requires at least two orthogonal virus reduction steps to comply with regulatory pathogen safety risk mitigation guidelines. Solvent detergent viral inactivation (SD VI) has historically been effectively utilized for the inactivation of lipid-enveloped viruses from plasma derived products, yet only recently have process development activities focused on implementation of SD VI in single use systems. The adoption of single-use technologies for bioproduction continues to progress, driven predominantly by the benefits of operational flexibility, speed of implementation, lower capital investment, and elimination of cross contamination concerns. However, the …

Single-Use Capacitance Measurement Of Cell Viability, Dan Kopec, Stuart Tindal, Marcus Weise, Sven Grob, Henry Weichert, Thomas Schnitzler, Manfred Biselli, Mario Becker

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

The biomanufacture of complex biologics continues to expand and develop with no end foreseen and in order to build on the industries rapid adoption of single use technology an ethos of what is measured and controlled in stainless steel can be, and should be transferable to plastic. The realization of this continues to be a challenge but one of the key critical parameters in cell cultivation; in-situ viable biomass is now quantifiable using single use capacitance measurement. A single-use sensor patch is fully integrated in a 5L Rocking Motion Bioreactor bag, gamma sterilized and applied to CHO DG44 and Hybridoma …

Material Science Enabled Quality By Design: The New Standard In Sus Development, Christian Julien

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

There is a growing interest from the end-user community in creating supply chain transparency as it pertains to critical raw materials used within their pharmaceutical manufacturing processes. Single-use systems contain mainly polymeric materials, often in diverse combinations drawing from a wide range of thermoplastic and elastomeric resins. Continued adoption of single-use assemblies into the entire spectrum of unit processing and production operations has led to increased scrutiny thereof and consequently many end-users have strengthened their technical diligence teams to include material scientists. These SME’s are tasked with enhancing the understanding of the materials of construction utilized in single-use systems. Their …

Adopting A Fully Single-Use Process To Improve Speed To Clinic: A Leachables Case Study, Jessica Shea, Elizabeth Goodrich, Ross Acucena, Paul Killian, Janmeet Anant

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

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 …

Interchangeability Of Sus - Functional Specification And Materials Of Construction, Chris Smalley

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Functionally equivalent substitution of an alternate supplier of a Single Use Component for an existing Single Use design provides a business contingency, or a process improvement, with equivalent performance and product quality. As an end-user, we are responsible for assessing the quality requirements to assure that the specifications are being met. This presentation will discuss the need for standards for designs and points to consider when implementing a Single Use Strategy that incorporates the potential for interchangeability

Assessing Cell Lines For Cell Growth Assays As Alternative For Existing Cytotoxicity Assays, Eike Jurkiewicz, Alexander Tappe

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Currently, biocompatibility assays based on USP Class VI or ISO 10993 standards are known to be unsuitable for evaluating the impact of SU technologies on biopharmaceutical processes. Although growth inhibiting substances like bis(2,4-di-tert-butylphenyl)phosphate (bDtBPP) have been present in raw materials of plastics, this specific substance has not been identified until recently (Hammond et al,. 2013) due to the insensitivity of commonly used cell lines. Up to today, the majority of plastic materials used in single-use containers for cell cultivation or media applications do not have the level of raw material control which is needed to ensure …

Acomdim Applied To Evaluate Gamma Irradiation Impact On Multilayer Pe Based Films, Fanny Gaston, Sylvain Marque, Nathalie Dupuy, Samuel Dorey

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

The use of single-use systems is becoming increasingly common in the biopharmaceutical and biotechnology industries. These systems are manufactured from polymers such as polyethylene (PE) and ethylene vinyl acetate (EVA). For their future applications, these devices are sterilized by γ-irradiation with a dose between 25 and 45kGy. C. Artandi and W.V. Winkle[i] determined that 25kGy is the dose to be at 40% above the minimum needed to kill the most resistant microorganisms. The purpose of this study is to understand what happens on the surface of polymers after γ-sterilization. Optical spectroscopy are of great interest for chemical analysis and …

Engineering Characterisation Of A Rocked-Bag Bioreactor To Evaluate Key Effectors Of Successful Mammalian Cell Culture, Douglas Marsh, Matthew Osborne, Martina Micheletti, Gary Lye

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Engineering characterisation is essential for efficient and knowledge-led process development in biomanufacturing and to underpin the adoption of single-use technologies [1]. Despite diverse applications of rocked bag bioreactors, there is currently little understanding of the fundamental determinants of fluid mixing and mass transfer, which are important for mammalian cell culture.

In this work, a flexible single-use rocked bag bioreactor system has been fully evaluated in terms of volumetric oxygen mass transfer coefficient (k_La), CO₂ stripping rate and liquid phase mixing time (t_m) at 10, 20 and 50L scale. Five inputs were identified as potentially affecting …

Evaluation Of Sweeping As An Alternative To Sparging In A Rectangular Single-Use Bioreactor, Jean Francois Hamel

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

The development of single use bioreactors (SUBs) has led to the exploration of novel reactor geometries such as the rectangular, single-use bioreactor (Nucleo™ Pad Reactor, PALL Corporation). Sparging is currently used as the main method of providing oxygen to cells in both the traditional and rectangular bioreactor. However, cell damage from bubble bursting and foam entrapment makes sparging a poor way of providing oxygen for growing cells that are sensitive to these shear inducing effects, such as stem cells. An alternative aeration method, sweeping, provides O₂ by diffusion through liquid air interface, minimizing shear stress damage to cells arising …

Sensors For Continuous Monitoring Of Disposable Bioreactors, Kenneth Reardon

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Continuous monitoring of biotechnological processes is important for control and optimization of quality and productivity. Typically, samples must be removed from the cultivation and analyzed in a laboratory to determine the concentrations of substrates and products. These time-delayed data cannot be used for real-time process control. The Process Analytical Technology initiative of the FDA supports the use of on-line measurement techniques for process development, production, and quality. Single-use systems are increasingly used in biotechnological applications, and face at least as many challenges as their stainless steel counterparts for process monitoring because the range of available sensors is limited to temperature, …

Single-Use Bag Extractable Case Study: Lessons Learned, Jayanthi Grebin, Mike Johnson

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

A supplier's ability to furnish extractable data from single-use components and systems to end users is of extreme importance. The data allows an end user to effectively assess the risk these extractables have on process fluids and final drug products. The protocols followed to produce such data are currently a point of discussion between suppliers and end users. In this case study, single-use bags were subjected to a portion of the BioPhorum Operations Group (BPOG) extractable protocol. This poster documents the testing process and lessons learned when executing such an evaluation, along with suggestions to improve the uniformity of the …

Microbial Applications Of Single Use Bioreactor Systems, James Rutley, Gary Lye

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

This work describes the ongoing assessment of single use bioreactors (SUBs) and their suitability for culturing microbial expression systems. Microbial expression systems are responsible for significant revenues as part of the biotechnology industry, in fact, their projected value for 2016 is in excess of $250 billion [1]. This promotes further interest in the area, with novel expression systems and pathways constantly under construction for the generation of bulk and high value chemicals.

The advantages offered by single use bioreactor technology has lead to increased adoption in many areas of bioprocessing [2], and microbial fermentation is no different. Applications include simple …

Challenges Of Scale Down Model For Disposable Bioreactors: Case Studies On Growth & Product Quality Impacts, Jincai Li, Guangming Zhang, Huilin Zhu

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Despite wide-spread use of disposable bioreactors, there is a lack of well-established scale-down model for larger scale SUBs. Here we report a case of NS0 cell culture process transfer from 2000L stainless steel bioreactor (SST) to 2000L disposable bioreactor (SUB). Initial attempts in trying to grow the NS0 cells in the small scale 2D bags yielded non-satisfactory results, as growth was impacted by bag material type as well as by suppliers of the same bag material type. However, 3D bags of 50L and above proved to be supportive of the NS0 cell line growth.

Even for cell lines that do …

Conversion Of Polymers – Properties And Processing, Phillip Jordan

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

The use of polymeric materials for manufacturing single-use disposable devices in biopharmaceutical processes to replace multi-use stainless steel systems is a steady trend.

A general overview on thermoplastic polymers is given: Commodity Materials, Engineering Plastics and High-Performance Plastics, with the focus on polymers frequently used in biotechnology applications.

This range of material covers rigid materials for components like connectors, filling needles, filters, etc., but also flexible materials for bags and tubing.

Thermoplastic Elastomeric materials (TPE) and Silicone are used for tubing applications. TPE materials can be seen as some kind of hybrid between thermoplastic and elastomeric polymers.

The characteristic conversion …

What's In The Bag?, Rebecca Olsen, Diane Hahm, Jing Li

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

As the bioprocessing industry continues to shift towards single-use systems, there is an increasing need for polymers with improved functionality. This article includes a review of the chemistry and functionality of various polymeric families, and a description of various tools that can be useful. Polyethylene resins are largely used in a variety of single use applications. This paper describes polyethylene chemistry in more detail so that single use system manufacturers and biopharmaceuticals manufacturers can optimize the use of polyethylene polymers in a particular application. The process in which the families of polyethylene polymer resins are manufactured dictates the degree of …

The Practical Application Of Bpog E&L Protocols To Viral Clearance Filters, Paul Killian, Thomas Stone, Jessica Shea

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

Regulatory guidance advocates virus control at various stages of the drug manufacturing process and directs that you test the capacity of the process to remove or inactivate virus. Patient safety concerns require you to determine what impurities may be added by the virus control steps you implement. While the application of a standardized approach to identifying and quantifying the extractables from these steps has benefits when making comparisons, choices have to be made when developing the protocol that take into account the characteristics of the clearance device and use conditions This presentation will illustrate the practical implementation of standardized extractables …