Engineering Commons^™

Spouting Behavior Of Binary Mixtures Of Spherical And Cylindrical Particles, Douglas W. Marshall, Nicholas W. Eimers, Nathan A. Yergenson, David C. Drown

Fluidization XV

Circulation of particles in a spouted bed is influenced by the aspect ratio of the particles. Circulation of high-aspect particles, such as grains, is similar to bulk flow of powders within a hopper, while circulation of spherical particles is better represented by funnel flow.

Deposition of high quality and high integrity coatings on nuclear fuel particles requires good circulation of the fuel particles to ensure a narrow distribution of coating thicknesses and uniform coating properties. Spouting studies were conducted with using air at room temperature in a 2-D model of a nuclear fuel particle coater to investigate the influence of …

Heat Transfer Challenge And Design Evaluation For A Multi-Stage Temperature Swing Adsorption (Tsa) Process, Gerhard Hofer, Tobias Pröll, Johannes Fuchs, Gerhard Schöny

Fluidization XV

Functionalized solid amine-based temperature swing adsorption (TSA) processes have recently been proposed as a potential way to reduce the energy-penalty of post-combustion carbon capture processes (1). If TSA is to be carried out at large scale and with high energy-efficiency, continuously operated counter-current contactors are required for thermodynamic reasons. This could, generally, be achieved by using moving bed contactors. However, the heat exchange requirement of TSA is significant and heat transfer is poor in fixed and moving beds. Therefore, multi-stage fluidized bed contactors with counter-current flow of solids and gas phase and immersed heat exchange surfaces may solve the heat …

Hydrodynamics Of Compartmented Fluidized Beds For Concentrated Solar Power Applications, Roberto Solimene, Riccardo Chirone, Andrea Paulillo, Piero Salatino

Fluidization XV

Application of fluidized beds to collection and thermal storage of solar radiation is beneficial in Concentrated Solar Power (CSP) systems thanks to their well-known inherently good thermal performances. Non-conventional design and operation of fluidized beds based on uneven or unsteady (pulsed) fluidization (1), may further enhance their thermal performances improving the potential for applications in the very demanding CSP systems. Dense gas-fluidized beds have the potential to effectively accomplish three complementary tasks: the collection of incident solar radiation; the heat transfer of the incident power to immersed tube bundles of high-efficiency steam and/or organic Rankine cycles (ORC) and the thermal …

Attrition Rate Of Iron Ore In The Gas-Solid Fluidized Beds With The Wide Size Distribution, Dong-Hyun Kang, Chang Kuk Ko, Dong Hyun Lee

Fluidization XV

The effects of superficial gas velocity (U_g = 1.25 – 3.00 m/s) and distributor hole size (8.0 – 12.4 mm) on the attrition rate of iron ore in a gas-solid fluidized bed with 0.076 m ID ´ 3.7 m height with or without circulation were investigated. The particle density and the Sauter mean diameter of fresh iron ore were 3,705 kg/m³ and 357 m, respectively. When the kinetic energy rate from the orifice was equal or greater than 180 J/s, the trend of attrition rate could be determined. The attrition rate was determined by measuring the fractional mass …

Carbon Dioxide Recovery By Means Of Tsa In A Sound Assisted Fluidized Bed Of Fine Activated Carbon, Raganati Federica, Ammendola P

Fluidization XV

A large decrease in CO₂ emissions through capturing and separation will be required to keep greenhouse gases at tolerable levels (1). Though several CO₂ capture technologies have been proposed, temperature swing adsorption (TSA), consisting in adsorbing the CO₂ on a solid material and, then, inducing the sorbent regeneration and CO₂ recovery by a temperature increase and gas purge, has the potential to become one of the leading techniques by complementing or replacing the current absorption technology due to its low energy requirement (2). With reference to the sorbent, great attention is focused on fine powders (3). …

Predicting Gas-Flow Distribution In Pilot-Scale Fluidized Beds Using Cfd Simulations, Akhilesh Bakshi, C. Altantzis, A. F. Ghoniem, L. R. Glicksman

Fluidization XV

Bubbling fluidized beds are used extensively in energy and chemical industries because of their excellent heat and mass transfer characteristics. Recently, CFD has been identified as a useful tool for predicting reactor performance, but application to large scales continues to be challenging because of limitations on computational resources. Given that the hydrodynamics can largely be characterized by bubbles rising through the bed, a more feasible approach for investigating large-scale reactors is to quantify bubble dynamics and specifically, gas distribution in different phases- visible bubble flow, bubble-through flow and dense-phase flow.

In this study, 3D CFD simulations of bubbling fluidized beds …

Novel Method To Meassure Fine Particle Circulation Rates In Draft Tube Conical Spouted Beds, Idoia Estiati, Haritz Altzibar, Mikel Tellabide, Martin Olazar

Fluidization XV

The spouted bed regime is an alternative contact method to fixed and fluidized beds. The ratio between the inlet diameter and particle diameter limits the scaling-up of spouted beds (the inlet diameter should be smaller than 20-30 times the particle diameter). The insertion of a draft tube is the way of overcoming this limitation.

Particle cycle time is defined as the time the particle takes to travel from the top of the annulus downwards and back again to its starting point. Since the proportion of time spent by a particle in the spout is insignificant compared with that spent in …

Modelling Study Of Two Chemical Looping Reforming Reactor Configurations: Looping Vs. Switching, Joana F. Morgado, Schalk Cloete, John Morud, Thomas Gurker, Shahriar Amini

Fluidization XV

Autothermal Chemical Looping Reforming (CLR) is a promising technology for hydrogen production with integrated CO₂ separation. Conventional CLR employs two fluidized beds (fuel and air reactors) with an oxygen carrier circulating between them. In this way, CLR supplies heat to the endothermic reforming reaction while avoiding fuel/nitrogen mixing. This configuration can achieve steady operation and low gas leakages between reactors, but has some drawbacks. The complex interconnected configuration is challenging to scale up, especially under the pressurized conditions required for high process efficiency. Moreover, the external circulation of particles through cyclones and loop seals increases reactor costs and imposes …

Characterization Of Fuel Segregation In A Fluidized Bed By Magnetic Particle Tracking, David Pallarès, Anna Köhler, Alexander Rasch, Filip Johnsson

Fluidization XV

A Magnetic Particle Tracking (MPT) system is used to experimentally characterize the vertical distribution of fuel particles in a fluidized bed. The measurements are carried out in a fluid-dynamically down-scaled cold unit resembling a 1×1 m2 boiler operating at 850 °C. The MPT system yields spatial and temporal information of the tracer particle with accuracies in the order of 10-3 m and 10-3 s, respectively. The MPT system is used to study how fuel segregation is influenced by physical properties of the fuel (sizes and densities representing different fuel types) and operational conditions, such as fluidization velocity, bed height and …

Conference Program (Gpe 2016), Jamal Chaouki, Franco Berruti, Xiaotao (Tony) Bi, Ray Cocco

Fluidization XV

No abstract provided.

Fed-Batch Process Development Using Metabolically Efficient Cho Cells, Cecile Toussaint

Cell Culture Engineering XV

The development and application of fed-batch strategies have greatly contributed to the improvement of the productivity of cell culture processes. However, in such processes, lactate and ammonia accumulation in the culture medium over time can have detrimental impacts on cell growth and product quality. Continuous cell lines typically exhibit an inefficient metabolism whereby most of the pyruvate derived from glucose is diverted to lactate and only a small percentage is incorporated into the TCA cycle. Thus, further process improvements can be expected by combining the application of rational fed-batch strategies with targeted metabolic engineering of cells to reduce waste metabolite …

Deepening Knowledge On Cho Cells Metabolism Using Multiple Tracer Substrates, Ana Teixeira, Manuel Carrondo

Cell Culture Engineering XV

Optimization of culture conditions through trial-and-error has led to much of the past progress on animal cell culture. Today, a renewed account of the underlying biologies through a “systems” view is providing a deeper knowledge of cellular physiology and laying the basis to reengineer desirable states of further increased culture performance. In this work, we took a comprehensive view of cellular metabolism by extensively profiling extracellular and intracellular metabolomes after ¹³C‑label supplementation in parallel labelling cultures of [1,2-¹³C]Glucose, [U-¹³C,¹⁵N]Asn, [U-¹³C,¹⁵N]Ser and [1-¹³C]Pyruvate. This integrative approach was used …

A Holistic Approach To Facility Protection From Adventitious Agents – A Case Study, Matthew Osborne, Ronan Kelly, Ann Maria Mccrohan, Marie Murphy

Cell Culture Engineering XV

The Eli Lilly biologics manufacturing facility in Kinsale, Ireland has been operational since 2010 with a 100% cell culture contamination control success rate. The presentation will review the holistic approach to facility protection from adventitious agents that underpins this success including:

• The risk assessment approach to points of entry and management via detectability and control measures
• The approach to personnel training that considers human factors, increased vigilance and event simulations following strategies that are used in chemical synthesis process safety

The presentation will then focus on control of adventitious virus. The talk will briefly comment on the early warning measures …

Building Qbd Frameworks Retrospectively For Commercial Products And The Use Of Scale-Down Model Qualification Strategies To Support Continuous Improvement, Jose Menezes, Francisca Gouveia, Pedro Felizardo

Cell Culture Engineering XV

A science-based justification for many continuous improvement and post-approval submissions on ‘legacy products’ (i.e., commercial products developed without a QbD rationale) requires that some of those missing QbD elements in the original filing, be defined retrospectively.

Here we present a new proposal to achieve that, that is general and very capable of supporting the transition of ‘legacy’ non-QbD into quasi-QbD filings. It is based on (1) a formal criticality analysis to the current process version to support the choice of quality attributes and process parameters in the filing, and identification of additional ones that may impact the quality target-product profile …

Separation Of Igg Glycoforms For Biosimilars Development Using Fc Gamma Receptors As Affinity-Based Chromatography Ligands, Austin Boesch

Cell Culture Engineering XV

Abstract: In biosimilars development, matching the glycosylation pattern of the innovator molecule can be a significant challenge in cell culture development. Here we present a novel downstream method using Fc gamma receptors as chromatography affinity ligands to separate monoclonal antibody glycoforms. Additional data is presented highlighting glycoform potentiation of effector functions such as antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis which can be important mechanisms of action to consider in biosimilars development

Nanofiltration As An Effective Means To Prevent Virus Contamination Of Cell Culture Processes, Christina Carbrello

Cell Culture Engineering XV

While cell culture media is typically sterile filtered to remove bacteria and mycoplasma, many bioreactors remain unprotected from viral contamination. As awareness of the risk for viral contamination of cell culture processes grows, manufacturers are seeking virus barrier technologies to remove or inactivate viruses. Filtration is an ideal technology in terms of familiarity, ease of use and scalability. However, hurdles to its implementation remain, most notably, the high cost and large footprints required to implement filters designed for the downstream process and the concern that the small pore size of virus filters could impact cell culture performance. A filter specifically …

Improvement Of Cho Specific Productivity Using Amino Acid Derivatives, Aline Zimmer, Caroline Hecklau, Sascha Pering, Alisa Schnellbaecher, Thomas Eichhorn

Cell Culture Engineering XV

Industrial fed-batch cultivation of mammalian cells is used for the production of therapeutic proteins such as monoclonal antibodies. Besides medium ensuring initial growth, feeding is necessary to improve growth, viability and antibody production. Amino acids are key elements for the cellular metabolism but also for the quality of the recombinant protein produced. Several studies have already demonstrated the link between amino acid availability and sequence variants [1-3] or specific modifications like trisulfide bonds [4]. To avoid such modifications, the chemical modification of amino acids is an interesting alternative to modulate their overall solubility [5], …

Development Of An Enriched Cho Feed Media For Quality Therapeutic Antibody Production From High Performing Clones, David Ho

Cell Culture Engineering XV

The use of feed media during protein production is essential to sustain viable cell densities and facilitate high productivity for modern high performing clones. However, the introduction of feed media can also have a significant impact on product quality attributes, such as glycosylation for therapeutic antibodies, which can ultimately impact therapeutic efficacy. For certain antibodies, the absence of fucose on the N-glycan at Asparagine 297 of the Fc domain has been shown to significantly increase binding to the FcgammaRIII receptor on NK cells and results in increased ADCC activity. Here we have incorporated product quality analysis including N-glycan analysis into …

Using Definitive Screening Design To Effectively Assess The Combinatorial Impacts Of Media Supplements On Monoclonal Antibody Production In Mammalian Cells, Aaron Chen

Cell Culture Engineering XV

Process understanding is an essential component of QbD principles. Cell culture media composition can impact many process performance parameters such as viable cell density, product yield and product quality attributes. In order to enhance process understanding and exert better process control, numerous media supplements have previously been tested. However, their combinatorial effects on process and product quality have largely been unexplored. This is partially due to the resource and efforts required to execute relatively large size of experiment and data analysis.

Recent advancement in modern design of experiment methodology has alleviated this issue. The definitive screening design (DSD) allows for …

Adapting An In-Licensed/Acquired Cell Culture Process To Platform Conditions, Raghunath Shivappa

Cell Culture Engineering XV

The use of platform materials and processes for new products help streamline process development, enhance predictability and efficiency during manufacturing, and simplify the raw materials supply chain. Although common, adaptation of cell expression systems can require a significant amount of process development work to achieve desired product yield, quality, and comparability. Here, we present a progressive development approach for evaluating platform media and feeds for an acquired cell culture process.

A CHO-DG 44 cell line expressing a novel antibody product was procured through acquisition. The original non-platform media and fed-batch process yielded titers of approximately 3 g/L. A study was …

Antibody Production With Site-Specific Non-Natural Amino Acid Incorporation For Generation Of Antibody Drug Conjugates, Alyssa Powell

Cell Culture Engineering XV

Ambrx’s mammalian expression platform (EuCODE) enables site-specific incorporation of non-natural amino acids into antibodies. This EuCODE technology provides a means for stable payload linkages at defined sites and with a defined DAR of 2 for antibody drug conjugate (ADC) generation. While the ability to control the DAR and payload site can provide an advantage to an ADC, the incorporation of the non-natural amino acid into the antibody heavy chain introduces a unique challenge for antibody production.

To achieve high production of monoclonal antibodies (mAbs) containing non-natural amino acids, we engineered a CHO-K1 cell line which stably contains Ambrx’s patented tRNA …

Development Of A Chemically Defined Media And A Chemically Defined Feeding Strategy For Extended Growth And Enhanced Productivity In Cho-K1 And Cho Dg44 Cultures, Sagar Kokal, Kyle Liu, John Menton

Cell Culture Engineering XV

The biopharmaceutical industry utilizes Chinese Hamster Ovary (CHO) cells for the production of recombinant proteins. Due to regulatory restrictions, there has been much focus on using serum-free culture processes including chemically defined (CD) media and feed system. A great deal of time and cost has been invested by biopharmaceutical companies for the development of an optimal CD media. Currently, the fed-batch process is the most dominant method for the large cultivation of CHO cells. In response to these industry requirements, Kerry has developed a complete system including a CD media called Sheff-CHO CD complete and a CD feed system named …

Evaluating Sugar-Based Detergents As A Potential Alternative To Poloxamer Bubble Protectant, Jessica Wuu, Jayanthi Lakkyreddy, Steven Meier

Cell Culture Engineering XV

Poloxamer 188 (commonly referred to as Pluronic® F-68) is a commonly-used medium component which protects against bubble-induced shear in mammalian cell cultures. Recently, concerns have arisen due to limited sourcing and variable performance of this critical component. In 2008, Hu, et al.¹ proposed the use of sugar-based detergents – specifically, maltopyranosides and glucopyranosides – as potential bubble protectants based on their rheological properties and relatively low toxicity to cells. This work explores the feasibility of these detergents using commercially relevant CHO cell lines and media. Detergent candidates were evaluated based on their tolerance by cells, stability in cell culture …

Adaptation Of Cho Metabolism To Long Term Phosphate Limitation, Mugdha Gadgil

Cell Culture Engineering XV

Phosphate is an important component of abundant intracellular molecules like RNA, is a critical component of energy metabolism and is involved in substrate phosphorylation in cellular metabolism and signaling. Inorganic phosphate (P_i) is transported through specialized transporter proteins and limitation in the availability of phosphate in the growth medium limits cell growth. P_i concentrations have been reported to regulate the rates of aerobic glycolysis and oxygen uptake. Thus it is expected that cellular metabolism will adapt to long term phosphate restriction. Since complete phosphate deprivation will result in lack of growth, such an adaptation needs to be …

Impact Of Poloxamer 188 Variability On Biologics Manufacturing: Mitigations And Causal Investigation, Salim Charaniya

Cell Culture Engineering XV

Poloxamer 188 (P188), a water-soluble, synthetic copolymer with hydrophilic as well as lipophilic properties, is widely used as a cell protectant for bubble-induced shear during cultivation of suspension-adapted mammalian cells for large-scale production of protein-based therapeutics. Poor cell growth and lower product yield has been recently observed at commercial scales in more than one Chinese hamster ovary-based manufacturing process within Roche network when specific lots of P188 were used. An investigation is being conducted to unearth the root cause of “poor-performing” P188 lots and to identify means to mitigate the product supply risks. P188 raw material is currently sourced from …

A Host Cell Protein That May Impact Polysorbate Degradataion, Kelvin Lee

Cell Culture Engineering XV

There is growing interest and concern related to host cell proteins. Current issues with host cell proteins include questions related to the various available analytical approaches and the sensitivity of those approaches as well as issues with purification strategies and the impact of host cell proteins downstream. The availability of a sequenced genome enables new opportunities to address problematic host cell proteins via cell line engineering. We studied difficult to remove host cell proteins in CHO cells with the goal of identifying host cell proteins that may be product associated with a number of monoclonal antibodies, that may copurify with …

The Differential Polarizability Of Cho Cells Can Be Used To Monitor Changes In Metabolism, Katrin Braasch

Cell Culture Engineering XV

The continuous monitoring of cell growth and viability is an integral part of biopharmaceutical production. Measurements of changes in the polarizability of individual cells can identify early emerging sub-populations of apoptotic cells in a dielectrophoretic (DEP) cytometer designed at the University of Manitoba. In this instrument the trajectory of individual cells was tracked as they passed through a bank of electrodes (sensitivity: 0.1 µm; rate: 5 cells per second) designed to differentially perturb the cells according to their polarizability. This perturbation was recorded as a force index (FI), which was related to the electrical displacement of the cells. Using this …

Development Of Antibody Detection Methods For Active Product At The Cell Culture Stage, Gregory Walsh, Matthew Schraut, Jon Barricelli, Lada Laenen

Cell Culture Engineering XV

Development of new analytical methods to characterize the manufacturing process became necessary when the site wanted to implement process improvements to increase output from the bioreactor. The bioreactor process is monitored and controlled for dissolved oxygen content, glucose concentration, temperature, etc. and samples of the harvest are analyzed daily using an endpoint assay and a total protein method. Neither of the two methods can detect the total amount of product at the harvest stage; the endpoint assay will only detect active product and the total protein method will not discriminate between the cellular proteins and the active product. We have …

Development And Application Of Glycosyltransferases For In Vitro Glycoengineering, Alfred Engel

Cell Culture Engineering XV

Glycosylation is an important posttranslational modification of proteins influencing protein folding, stability and regulation of the biological activity, e.g. IgG featuring terminal sialic acids were shown to suppress inflammatory response (ADCC) and increase serum half-life. Fully galactosylated structures seem to improve binding to the complement system (CDC).

Despite all efforts in cell-line engineering and process optimization to increase glycosylation homogeneity, today therapeutic proteins still show a heterogeneous glycosylation pattern, with large variations between bioprocesses and even from batch to batch. The use of glycosyltransferases for enzymatic synthesis of well-defined glycan structures will become an essential tool – at least in …

Comparison Of Commercial Cho Cell Media Formulations Using Material-Oriented Recurrent Spectral Libraries, Kelly Telu

Cell Culture Engineering XV

Chinese hamster ovary (CHO) cells are commonly used for the production of biological therapeutics. Metabolic profiles of media components can be used to monitor process variability and look for markers that discriminate between batches of product. Currently, there exists no database of CHO media components or a method for the comparison of commercial media formulations. We are creating material-oriented libraries of CHO media components that include LC-MS/MS and GC-MS data. The libraries represent all metabolites that can be detected by LC-MS/MS and GC-MS and consist of recurrent spectra that cover all known fragmentation conditions and precursors. Recurrent spectra occur repeatedly …