Engineering Commons^™

Mir-Catch Identifies Biologically Active Mirna Regulators Of The Pro-Survival Gene Xiap, In Chinese Hamster Ovary Cells, Paul S. Kelly, Alan Griffith, Sebastian Vencken, Nga T. Lao, Martin Clynes, Niall Barron

Cell Culture Engineering XVI

Genetic engineering of mammalian cells, in particular Chinese hamster ovary (CHO) cells, is of critical interest to the biopharmaceutical industry as a means to further boost the yields of therapeutic proteins. Complimentary to already in place advanced bioprocesses, stable overexpression of the pro-survival X-linked inhibitor of apoptosis (XIAP) is one example of the successful manipulation of CHO cell genetics resulting in prolonged culture survival, ultimately increasing recombinant protein productivity. However, saturation or burdening of the cells translational machinery can occur in instances of forced expression of a trans-gene thereby achieving the anticipated cellular phenotype without the associated improvement in productivity. …

Aligned Carbon Nanotube Arrays Bonded To Solid Graphite Substrates: Thermal Analysis For Future Device Cooling Applications, Betty T. Quinton, Levi Elston, James D. Scofield, Sharmila M. Mukhopadhyay

Mechanical and Materials Engineering Faculty Publications

Carbon nanotubes (CNTs) are known for high thermal conductivity and have potential use as nano-radiators or heat exchangers. This paper focuses on the thermal performance of carpet-like arrays of vertically aligned CNTs on solid graphite substrates with the idea of investigating their behavior as a function of carpet dimensions and predicting their performance as thermal interface material (TIM) for electronic device cooling. Vertically aligned CNTs were grown on highly oriented pyrolytic graphite (HOPG) substrate, which creates a robust and durable all-carbon hierarchical structure. The multi-layer thermal analysis approach using Netzsch laser flash analysis system was used to evaluate their performance …

Irradiation Experiment For Living Insect-Based Radiological Dispersal Device, Braden Goddard, Walid Metwally, Ahmad Ababneh

International Journal of Nuclear Security

People generally associate fear with “nuclear”, “radioactive” and “insects”. It is speculated that a release of radioactive living insects would instill more fear into the general public than a “traditional” style radiological dispersion device (RDD). This paper evaluates the potential threat of an insect-based RDD using experimental data. The results of this project found that insect-based RDDs are an insignificant threat due to the challenges in making insects radioactive enough to pose any danger to humans without killing the insects.

Driver Responses To Graphic-Aided Portable Changeable Message Signs In Highway Work Zones, Yilei Huang, Yong Bai

Civil and Environmental Engineering Faculty Research and Publications

Portable changeable message signs (PCMSs) have been employed in highway work zones as temporary traffic control devices. Various studies showed that adding graphics to PCMS messages can provide advantages to traditional text messages, such as increasing legibility and improving the understanding of elderly drivers. This article synthesizes the findings of a two-phase research project aimed to investigate driver responses to graphic-aided PCMSs. Different text and graphic-aided PCMSs representing roadwork and flagger were set up in the upstream of highway work zones, and speed data of more than 2,700 vehicles were collected with a series of five speed sensors to determine …

Growth Of An Algae Polyculture And Dunaliella Tertiolecta In Oilfield Produced Water At A Range Of Salinities And Nutrient Concentrations For Biofuels Production., Thomas C. Hopkins

Civil Engineering ETDs

Growth of algae in saline produced waters (wastewater) from oil and gas production is of interest for potential production of algal biofuels, wastewater treatment and potential reuse or disposal. The objectives of this study were to determine the effects of salt concentrations and nitrogen source (ammonia or nitrate) and concentration on algal growth, lipid production, and nutrient removal to help determine suitability for growth on saline produced waters. Dunaliella tertiolecta (a marine algae) and a polyculture (previously discovered growing in PW) were grown on produced water with a range of salinities and varying nitrogen source and concentration, in a series …

Investigation Of The Use Of A Bio-Derived Solvent For Non-Solvent-Induced Phase Separation (Nips) Fabrication Of Polysulfone Membranes, Xiaobo Dong, Amna Al-Jumaily, Isabel Escobar

Chemical and Materials Engineering Faculty Publications

Organic solvents, such as N-methyl-2-pyrrolidone (NMP) and dimethylacetamide (DMAc), have been traditionally used to fabricate polymeric membranes. These solvents may have a negative impact on the environment and human health; therefore, using renewable solvents derived from biomass is of great interest to make membrane fabrication sustainable. Methyl-5-(dimethylamino)-2-methyl-5-oxopentanoate (Rhodiasolv PolarClean) is a bio-derived, biodegradable, nonflammable and nonvolatile solvent. Polysulfone is a commonly used polymer to fabricate membranes due to its thermal stability, strong mechanical strength and good chemical resistance. From cloud point curves, PolarClean showed potential to be a solvent for polysulfone. Membranes prepared with PolarClean were investigated in terms of …

Pedestrian Injury Severity Analysis In Motor Vehicle Crashes In Ohio, Majbah Uddin, Fahim Ahmed

Faculty Publications

According to the National Highway Traffic Safety Administration, 116 pedestrians were killed in motor vehicle crashes in Ohio in 2015. However, no study to date has analyzed crashes in Ohio in order to explore the factors contributing to the pedestrian injury severity resulting from motor vehicle crashes. This study fills this gap by investigating the crashes involving pedestrians exclusively in Ohio. This study uses the crash data from the Highway Safety Information System, from 2009 to 2013. The explanatory factors include the pedestrian, driver, vehicle, crash, and roadway characteristics. Both fixed- and random-parameters ordered probit models of injury severity (where …

Validation Of Cfd Simulations For Hypersonic Flow Over A Yawed Cone, Julian Cecil

McKelvey School of Engineering Graduate Student Theses & Dissertations

This study aims to numerically simulate the wind tunnel results for hypersonic flow over a circular cone of semi-apex angle of 10 degrees yawed from 0° to 20° using the commercial computational fluid dynamics software ANSYS Fluent. The ANSYS workbench is used to create the 10° semi-apex circular cone with a shock aligned structured mesh of 3.05 million cells surrounding the cone. Simulation boundary conditions for pressure and temperature in the far field correspond to Tracy’s wind tunnel experiment at Cal Tech. The six simulations cases are conducted for yaw angles of 0, 8, 12, 16, 20 and 24 degrees. …

Controlling Continuous High Cell Density Perfusion Culture With The Alternating Tangential Flow System In Real Time Using Radio-Frequency Impendance, Aditya Bhat

Cell Culture Engineering XVI

Many cGMP cell culture processes are now based on a continuous high cell density perfusion bioreactor system. Control of the feed or addition rates to maintain pseudo-steady-state conditions in these bioreactors can be especially challenging due to high and fluctuating cell concentrations that can rapidly change environmental conditions. With infrequent manual daily sampling based on offline cell counting, the control system can have too little information on which to base an appropriate decision to manipulate the process. Tight control of the perfusion or concentrate addition rate allows the bioreactor to be operated under the optimum conditions for maximum recombinant protein …

Scalable Lentiviral Vector Production Using Stable Producer Cell Lines In Perfusion Mode, Sven Ansorge, Aziza Manceur, Stéphane Lanthier, Sonia Tremblay, Anne-Marie Gélinas, July Dorion-Thibaudeau, Sophie Broussau, Julia Transfiguracion, Hafida Aomari, Rénald Gilbert

Cell Culture Engineering XVI

Lentiviral vectors (LVs) are becoming an important tool in gene and cell therapy and are being utilized in several clinical studies against genetic and acquired diseases, as well as in cancer therapies. To address the challenges linked to the generation of preclinical and clinical supply, the National Research Council Canada has developed packaging cell lines and stable producer cell lines for the production of LVs which can grow in suspension in serum-free media and produce LV in the 106 TU/ml range without optimization. We focus on the development of perfusion processes to both intensify the process and harvest produced LV …

Dxb11/Taut Host Cell Engineering Strategy Enabling The Establishment Of Strains Producing The Highest Yield Of Advanced Recycling Antibodies, Hisahiro Tabuchi, Masaki Yamaguchi, Tao Jialin, Hirokatsu Makitsubo

Cell Culture Engineering XVI

Innovation in monoclonal antibody (mAb) production for clinical use continues to be driven by cell engineering strategies to increase mAb yield and to control the complexity of advanced recycling antibodies (rcAbs). rcAbs offer significant advantages for efficacy by dissociating the antigens in endosomes and recycling free antibodies back to plasma. We were able to reduce the instability in an rcAb-producing CHO cell line whereby mAb titer during the stage of cell line development (CLD) was decreased and improve cell culture optimization. In this study, we used a chemically defined medium (CDM) and CDM-adapted transporter-overexpressing DXB11 host cells for CLD of …

Novel Transposase Tools For Cell-Line Engineering, Ferenc Boldog, Jeremy Minshull, Mark Welch, Kate Caves

Cell Culture Engineering XVI

ATUM has discovered, characterized, engineered and patented new transposases that work comparable to, or better than previously commercialized transposases. The technology is highly valuable for protein expression and genome engineering applications. It enables a specified sequence to behave as a transposon, a mobile genetic element, which can efficiently transpose between vectors and chromosomes via a “cut & paste” mechanism. The Leap-In Transposase® catalyzes the integration of a transposon containing your gene into TTAT sites in the target genome. During transposition, the Leap-In Transposase recognizes transposon-specific inverted terminal repeat sequences (ITRs) located on both ends of the transposon vector and moves …

Empowering Manufacturing Decisions Through Process Simulation Models, Tiffany Rau, Terence Davidovits, Rick Stock

Cell Culture Engineering XVI

The need for Clinical and Commercial manufacturing capacity continues to increase and at the same time there are “shortages” of production slots and time pressures. Whether one is a virtual, small or large company there are many considerations that go into the decision of how and where to manufacture the biopharmaceutical, cell therapy or vaccine. In this presentation, we explore approaches for using commercially available modeling tools, such as Super ProDesigner® and BioSolve™, to simulate the manufacturing of a “typical” monoclonal antibody product and a multivalent conjugate vaccine product. Through these hypothetical case studies, we will illustrate several important concepts …

Conference Program, Anne Skaja Robinson, Raghavan Venkat, Gene Schaefer

Cell Culture Engineering XVI

No abstract provided.

Impact Of Raw Materials On Sialyation For A Therapeutic Protein, Wei-Chien Hung, Mei Shao, Bridget Leslie, John Mavrianos, Paul Loray, Mei Shao

Cell Culture Engineering XVI

Total sialic acid content (TSAC) is a critical product quality attribute (CQA) for a therapeutic protein. Previous studies and results have suggested lot-to-lot variability of raw material, media powder and feed, have potential effects on total sialyation acid content (TSAC) at cell culture fluid (CCF) and harvested cell culture fluid (HCCF). Through searching literature, we found that potential components in media powder and feed such trace metal and vitamins can greatly modulate the sialyation acid level in vivo or in vitro. For instance, the sialic acid content was simultaneously increased in glycopeptides as well as gangliosides under lithium treatment^{1 …}

Demonstrating A Powerful Scale-Up Strategy For Biosimilar Mab In Single Use Systems Via Physicochemical And Functional Characterization, Serdar Alpan, Ozge Can, Deniz Baycin, Yigit Erdemgil, Zeynep Yildirim Keles, A. Emin Atik

Cell Culture Engineering XVI

Biosimilars have received a remarkable attention in the recent years. Due to the heterogeneity of biosimilar mAbs, they need to be well-characterized by various orthogonal techniques in order to identify their physicochemical and functional characteristics. Characterization of the post translational modifications, especially, glycosylation is vital to define the critical quality attributes (CQAs) which affect safety, efficacy and quality of drugs. In this study, we were able to manipulate the quality of the drug by using scale-up strategies for single use systems. By using ultra-performance liquid chromatography (UPLC) coupled to mass spectrometry (MS), we were able to demonstrate physicochemical similarities between …

The Journey From Tech Transfer To Bla Submission: Case Study Of A Ns0 Cell Culture Process From 2000l Stainless Steel Bioreactor To 2000l Disposable Bioreactor, Jincai Li

Cell Culture Engineering XVI

A case study of NS0 cell culture process transfer from 2000L stainless steel bioreactor (SST) to 2000L disposable bioreactor (SUB), and through to process validation and BLA submission is reported for production of an antibody therapeutics in this poster. Initial attempts in growing 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 different suppliers of the same bag material type. However, 3D bags of 50L and above proved to be supportive of the NS0 cell line growth. Process characterization (PC) and process validation (PV) efforts …

Identifying Hipk1 As A Target Of Mir-22-3p Enhancing Recombinant Protein Production From Hek 293 By Using Microarray And Htp Sirna Screen, Sarah Inwood, Eugen Buehler, Michael Betenbaugh, Madhu Lal, Joseph Shiloach

Cell Culture Engineering XVI

Enhancing protein production in mammalian cells is of interest in the biomedical field for a variety of reasons, including structural studies and antibody production. Using small non-protein coding RNA such as microRNA has recently been a promising method of increasing protein expression. A high throughput human microRNA screen in HEK 293 cells previously identified miRNA 22-3p as a promising candidate for increasing recombinant protein expression. This microRNA enhanced the expression of luciferase, two hard-to-express membrane proteins and a secreted hFc-fusion protein. In order to explore the mechanisms of this increase in protein production and to understand the intracellular events, we …

Towards Model-Based Bioprocess Characterization: A Mathematical Model Of Cell Cycle, Metabolism And Apoptosis Of Mab-Producing Mammalian Cells, António Lima Grilo, Athanasios Mantalaris

Cell Culture Engineering XVI

The biologics industry is changing and all players seek competitiveness through process optimization aiming at satisfying the market demand providing a compliant protein in as short time as possible. As blockbuster drugs experience patent expiry, several biosimilars are coming market and competition becomes fiercer. Unlike classical experiments-based optimization, bioprocess modeling is a rational, economically efficient and reliable alternative to deliver an optimized bioprocess. The biological aspects of a bioreactor are diverse and include cells growing and cycling, other cells dying, nutrients being consumed and by-products accumulating, energy production and usage and therapeutic proteins being produced. The understanding of the biology …

Impact Of S-Sulfocysteine On Fragments And Trisulfide Bond Linkages In Monoclonal Antibodies, Aline Zimmer, Ronja Seibel, Alisa Schnellbaecher, Susanne Bohl, Maria Wehsling, Sandra Maier, Anne Zeck

Cell Culture Engineering XVI

The quality of recombinant proteins such as monoclonal antibodies produced using Chinese hamster ovary cell-based mammalian systems is dependent on many factors, including cell line, process and cell culture media. Due to these factors, the generated product is heterogeneous and may have chemically induced modifications or post-translational modifications that affect antibody stability, functionality and, in some cases, patient safety. This study demonstrates that S-sulfocysteine, a cysteine derivative, can increase the antibody specific productivity in different cell lines cultivated with different processes while minimizing trisulfide linkages in generated mAbs, mainly between heavy and light chain. The supplementation of a cell culture …

Strategies To Improve Productivity Of Cho-S Cells Expressing An Anti-Tnfα Monoclonal Antibody With Biosimilar Potential, Ana M. Moro, Matheus Luchese, Lilian Tsuruta, Wagner Quintilio, Mariana Lopes Dos Santos

Cell Culture Engineering XVI

Tumor necrosis factor alpha (TNFα) is a proinflammatory cytokine that mediates the homeostasis of immune responses; its exacerbated production is associated with the pathogenesis of autoimmune and chronic inflammatory diseases. Anti-TNFα drugs have revolutionized the treatment of inflammatory conditions such as rheumatoid arthritis and Crohn’s disease. Currently, a worldwide race is on stage for the production of biosimilars moved by patent expiration of monoclonal antibodies (mAbs), such as anti-TNFα adalimumab. This project is based on the first development stage for an adalimumab biosimilar candidate with potential for national production through the generation of a productive and stable cell line and …

Off-Target At-Scale Scale Down Model Verification Of A Marketed Biopharmaceutical, Markus Emmler, Stefanie Menrad, Yosuke Watanabe

Cell Culture Engineering XVI

According to authority guidelines, process understanding is considered the key factor for a successful control strategy in submissions for biopharmaceutical production processes. This knowledge is usually gained in scale down model (SDM) experiments and therefore is subject to the ability of the SDM to reliably predict behavior of cell culture processes in manufacturing scale. In order to prove predictability, SDM runs at target conditions are typically compared to manufacturing scale target runs only. At the same time, predictability is also assumed within the whole design space described or even beyond as evaluated in process design studies. The proof at off-target …

Development Of A Platform Expression System Using Targeted Integration In Chinese Hamster Ovary Cells, Scott Bahr, Trissa Borgschulte, Michael Johns, Jeff Galligan

Cell Culture Engineering XVI

In recent years the biomanufacturing industry has seen significant improvements in recombinant protein production titers due to advancements in protein expression technologies as well as media, feed and manufacturing process development. However, the standard methods of recombinant cell line development have remained relatively unchanged. The majority of the biopharma industry introduces transgenes into Chinese Hamster Ovary (CHO) cells using mechanical or chemical transfection processes followed by metabolic or antibiotic selection of stable recombinant pools. Through this process, the transgene(s) are randomly integrated into the genome, often times resulting in significant heterogeneity within the stable pools. Individual recombinant CHO cells within …

Curation Of A Cho Dg44 Genome Scale Model And Application To Support Cell Culture Development Process, Cyrielle Calmels, Laetitia Malphettes, Mikael Rørdam Andersen

Cell Culture Engineering XVI

Genome scale models (GSM) have become a useful tool to connect different omics dataset into a single computational framework, thus giving a good overview of the flux distribution and metabolites interconnections in a specific environmental condition. A community genome-scale metabolic network reconstruction of Cricetulus griseus and cell line specific models have been recently developed 1. The main objectives with the use of the published CHO DG44 model were to enhance industrial bioprocess performance by suggesting genetic or metabolic targets, as well as strategies for medium optimization, and by bringing more fundamental knowledge about CHO cell metabolism. In a first step, …

The Relevance Of Cell Size In A Cho Fed Batch Process: Metabolic And Transcriptomic Characterization, Dirk Martens, Xiao Pan, Rene Wijffels, Ciska Dalm

Cell Culture Engineering XVI

Normally the growth profile of a CHO cell fed-batch process can be divided into a growth phase followed by a stationary (non-growth) phase and a cell death phase. In this study, using a commercially available media system in a 10 liter fed-batch reactor, the growth phase is followed by a phase where cell division stops but cell growth continues in the form of an increase in cell size (Figure 1). During the cell size increase (SI) phase, the average volume and dry weight per cell increase linearly with time up to nearly threefold. Cell cycle and transcriptome analysis show that …

Tools And Methods For Providing Assurance Of Clonality For Legacy Cell Lines, Paul Wu, Juhi Ohja, John Thrift, Taymar Hartman, Louise Almond, Jennitte Stevens, Sam Yaghmour, Christina Alves, David Shaw, Mike Laird, Robyn Emmins, Yuan Zhu, Ren Liu, Zhimei Du, Rolf Koehler, Thomas Jostock, Karin Anderson, Howard Clarke, Chris Campbell

Cell Culture Engineering XVI

Over the last several years demonstration of cell line clonality has been a topic of many industry and regulatory presentations and papers. Guidance has been provided by the regulatory authorities, especially the FDA, on a path forward for providing evidence of clonality with high probability. It has been recommended that two-rounds of limiting dilution cloning (LDC) at sufficiently low seeding densities (≤0.5 cells/well) provides sufficient evidence that a cell line is clonal. Furthermore, one-round of LDC may also suffice if supplemental data from a characterized FACS or plate-imaging workflow are also included in the package.

Cell lines generated by methods …

Probability Distributions In The Glass Failure Prediction Model, Samir Blanchet, H. Scott Norville, Stephen Morse

Michigan Tech Publications, Part 1

Glass, a brittle material, fractures under tensile stress acting over a time duration. Lateral loads, such as wind, acting on a simply supported rectangular glass lite, put one surface of the lite primarily into tension. ASTM E 1300 defines load resistance of glass as the uniform lateral loading acting over a duration of 3 seconds that is associated with a probability of breakage of 8 lites per 1000 at the first occurrence of the loading. To determine load resistance, the underlying window glass failure prediction model facilitates determination of a probability distribution of 3 second equivalent failure loads, P3. The …

Ftc Regulating Cybersecurity Post Wyndham: An International Common Law Comparison On The Impact Of Regulation Of Cybersecurity, Andrew Z. R. Smith

Georgia Journal of International & Comparative Law

No abstract provided.

Energy Modeling & Design Of Prototype Hydroponic Grow System, Emily Denise Trawick, William James Stinson, Alexander Charles Martin

Senior Honors Projects, 2010-2019

Energy and food security relies on innovations that spur sustainable ideologies. This project considers a novel approach to grow microgreens within a controlled environment in a manner that conserves water, minimizes environmental impacts from agriculture runoff, and enables successful agriculture in virtually any environment. The eQUEST® software package, an energy simulator, has been used to create a model of the “grow box” considered in this study. The dimensions were specified, and heating, cooling, and other loads were incorporated into the model which was used to estimate energy consumption. Real-time data were collected from sensors installed in the container and were …

Atmospheric Resiliency: Enhancing Environmental Air Quality Through Bio-Architecture, Francesca Boakye

Bachelor of Architecture Theses - 5th Year

The theory of purifying a city’s smog filled atmosphere by using green microalgae can be architecturally realized through a designed building envelope composed of multiple units of microalgae filled perforated panels. The nature of this project is bio-architectural. Precise execution of it relies on deep research in vehicular emission pollution in terms of molecular composition. This research will help identify the most prominent pollutant in the smog composition and match it to a species of algae that can completely process it during photosynthesis. In addition to air pollution research, research on algae typology, growth and environmental impact is also essential …