Engineering Commons^™

Incidence And Potential Implications Of Methylglyoxal In Industrial Cell Culture Revisited, Frank Chaplen

Cell Culture Engineering XV

Methylglyoxal is a toxic by-product of glycolysis and amino acid metabolism in mammalian systems. The major route for methylglyoxal detoxification is the glyoxalase pathway, which consists of the enzymes glyoxalase I (GLO1) and glyoxalase II (GLO2). A required co-factor for the glyoxalase pathway is reduced glutathione. Evidence suggests that GLOI and methylglyoxal have important roles to play in the signal pathways associated with oxidative stress and necrotic cell death¹. Previous work has demonstrated that growth conditions found in industrial cell culture have marked effects on endogenous methylglyoxal levels in Chinese hamster ovary (CHO)². Furthermore, decreased levels …

Anti-Oxidant Addition To Cd-Cho Media To Prevent Damage Induced By Uv Disinfection, Emma Dare

Cell Culture Engineering XV

Ultraviolet (UV) irradiation is an advantageous sterilization technique in the biopharmaceutical industry as it is capable of targeting non-enveloped viruses and smaller viruses that are difficult to remove via conventional filtration. Unfortunately, cell culture media contains many components that, upon exposure to light, undergo phototransformations that subsequently lead to poor cell growth. We have shown previously Chinese Hamster Ovary (CHO) cell culture performance was affected at fluences above 200 mJ/cm² due to the depletion of several key nutritional components (pyruvate, tryptophan, tyrosine, pyridoxine, pyridoxamine, thiamine) and the accumulation of reaction by-products (acetate, formate, sarcosine). Our objective was to determine …

Accelerated Bioprocess Characterization By Data Enrichment In Scale-Down Models, Viktor Konakovsky, Graham Mccreath, Jarka Glassey

Cell Culture Engineering XV

Scale-down models are often used for the definition of operating ranges in process development and validation and therefore data exploitation is an important task. Usually, the experimental scientists have very tight timelines and consequentially may often only perform the required routine of cleaning and sorting the data without more sophisticated analyses, even though it might improve data quality. This problem is more exacerbated by the rise of fully automated, miniaturized high-throughput equipment in up- and downstream process development where data processing automation is not just optional but a must. However, cleaned, sorted and time-aligned data alone do not guarantee a …

Conference Program, Kathy Chan

Cell Culture Engineering XV

No abstract provided.

Nmr-Based Design Of Chemically-Defined Protein-Free Feed Medium For The Cho Expression System, Marina Goldfeld

Cell Culture Engineering XV

Recombinant expression of antibodies and other biotherapeutics relies heavily on the use of CHO cell lines. The product expression levels in these cell lines are dependent on the feed medium, one of the most important components of the cell culture process. We have developed a systematic approach to designing a chemically-defined feed medium through collaboration between Merck’s Process Development and Assay Development groups. Historically, feed media have been derived from non-chemically defined sources, such as bovine serum and yeast hydrolysate. While these media support robust cell growth and productivity, their chemically-complex nature has hindered efforts to increase the cells’ productivity. …

Troubleshooting The Recover Of Mater Cell Bank For A Commerical Product, Mei Shao, Kishan Rao

Cell Culture Engineering XV

Poor recoverability was observed for the Master Cell Bank (MCB) during the preparation of Working Cell Bank for a commercial product. The presentation will describe the investigation process, root causes identified and corrective actions implemented. A new Working Cell Bank was successfully manufactured after implementing the corrective actions. Our strategy to test the WCB prior to commercial manufacturing will also be discussed. In addition, preventative actions were also implemented as a result of the event.

The Oxygen Binding Protein, Hemoxcell(R), Increases Cho Cell Growth And Extends Viability By Enhancing Oxygen Delivery, 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 …

Moving From A Bioreactor Scale-Up/Scale-Down Approach To A More Holistic Operational Design Space View, Gene Schaefer, Rick Yeager, Ted Deloggio

Cell Culture Engineering XV

Scale-up of bioreactor processes for biologic products has been an area of focus in the industry for over 50 years and the results have generally been successful through the application of a number of specific correlations across scales for mass transfer, mixing, etc. However, the ability to visualize and understand the bioreactor design space in terms of these parameters has generally been limited as has any general understanding of how close a given process comes to certain sensitivity limits at various scales. As part of a project which involved establishing scale-down models at two different sites with long-term objectives of …

Implementation Activities For A Chemically-Defined Media Platform To Minimize Media Variability Impact To Cell Culture Performance And Product Quality, Martin Gawlitzek, Brian Wong, Robert Shawley, Masaru Shiratori

Cell Culture Engineering XV

Chemically-defined media have been developed for CHO cell culture platform processes. During implementation of the media at multiple scales and sites globally, additional activities were required to seamlessly balance practical considerations with desired process and product quality performance. Three main focus areas were further investigated and optimized to enable the platform implementation: 1) media preparation, filtration, and post-filtration stability; 2) media and supplement viral barrier compatibility; and 3) and raw material variability considerations. Overall, the effort provided an outlined approach for media implementation in the global network including scale and site-specific case studies where novel options were required.

The data …

Challenges In The Development And Adaptation Of Platform Process To Existing Pipeline, Edmund Scarfo

Cell Culture Engineering XV

The development and use of platform processes have been adopted as an industry-wide strategy for enabling faster timelines for early clinical stages. However due to increasing product demands and the competitive landscape of antibody therapeutics in the industry, the development and use of a highly productive and economic upstream platform process has also become an adopted strategy. In keeping with industry standards, Takeda’s platform has evolved from upstream processes capable of achieving protein titers from 0.7-1.2 g/L to highly productive processes achieving titers in the range of 4.0-7.0 g/L. Unlike these earlier, low titer processes for the same molecules, the …

How To Select The Most Suitable Media For Your Cells, Marcella Dalm

Cell Culture Engineering XV

Many different media are available to culture CHO cells. Most are either growth supporting or productivity supporting. Ideally, the basal medium is a growth supporting medium, while the feed medium is supporting both growth and productivity. In this study two clones producing the same mAb are compared on their response to a range of basal media. Clone I is fast-growing and low-producing, while Clone II is slower-growing, but high-producing. A panel of eleven different basal media from different vendors was evaluated for growth, titer and metabolism.

An overview is given on the response of the cells to the different media. …

Evaluation Of Performance Enhancing Effects Of Supplementation With Complex Feed System And Supplements With Sheff-Cho Cd Complete Media In Cho-K1 And Cho Dg44 Cultures, John Menton, Kyle Liu, Sagar Kokal

Cell Culture Engineering XV

Biopharmaceutical companies routinely utilize feed systems for maximizing the yield of recombinant proteins in Chinese Hamster Ovary (CHO) cells. Synergies between plant-derived hydrolysates, yeast extracts and recombinant proteins have previously been demonstrated to enhance productivity in CHO fed-batch cultures. Based on this knowledge, Kerry has developed CHO specific ACF (animal component free) feed systems and supplements for improving the overall culture health and enhancing recombinant protein yields. The availability of protein-free (PF) versions of the feeds and supplements provides an option for production platforms with PF requirements. Kerry has developed Sheff-CHO CD complete a CD media designed to work synergistically …

An Addition Of Lithium Chloride Improves The Transient Gene Expression Yield In Cho Cells, Che Kim, Tae Kwang, Gyun Lee

Cell Culture Engineering XV

As hundreds of new recombinant therapeutic proteins enter into development each year, a cost-effective transient gene expression (TGE) system is required to assess these proteins at an early phase as a more rapid alternative to a stable gene expression system. While several strategies, such as genetic manipulation and environmental modification, have been attempted in TGE systems to improve the overall yield in Chinese hamster ovary (CHO) cells, chemical supplementation is often employed in industrial processes owing to its simplicity and effectiveness. Lithium chloride (LiCl), which is known to induce G2/M-phase cell cycle arrest, was found to be an attractive chemical …

Retrospective Implementation Of Quality By Design For Legacy Commercialized Enzyme Replacement Therapies, Anup Agarwal, Eric Hayduk

Cell Culture Engineering XV

Quality by Design (QbD) has been widely adopted by the pharmaceutical industry as a tool for transforming development, manufacture, and commercialization of drug products. QbD ensures that quality is built into a manufacturing process to consistently produce desired product. As per a FDA guidance¹, QbD should be employed at the product development stage to ensure manufacture of a product with predefined quality. Here we present a retrospective QbD approach employed at Shire to define an improved control strategy for a commercial process using risk-based process understanding and characterization. Key challenges and considerations of implementing a QbD strategy on …

Utilization Of Distributed Generation In Power System Peak Hour Load Shedding Reduction, Nandu Balachandran

LSU New Orleans Theses and Dissertations

An approach to utilize Distributed Generation (DG) to minimize the total load shedding by analyzing the power system in Transactive energy framework is proposed. An algorithm to optimize power system in forward and spot markets to maximize an electric utility’s profit by optimizing purchase of power from DG is developed. The proposed algorithm is a multi-objective optimization with the main objective to maximize a utility’s profit by minimizing overall cost of production, load shedding, and purchase of power from distributed generators. This work also proposes a method to price power in forward and spot markets using existing LMP techniques. Transactive …

Sensitivity Analysis Of Synchronous Generators For Real-Time Simulation, Sowmya Munukuntla

LSU New Orleans Theses and Dissertations

The purpose of this thesis is to validate generator models for dynamic studies of power systems using PSS/E (Power System Simulator for Engineering), EMTP (ElectroMagnetic Transient Program), and Hypersim. To thoroughly evaluate the behavior of a power system in the three specified software packages, it is necessary to have an accurate model for the power system, especially the generator which is of interest. The effect of generator modeling on system response under normal conditions and under faulted conditions is investigated in this work. A methodology based on sensitivity analysis of generator model parameters is proposed aiming to homogenize the behavior …

A Study Of Three Paradigms For Storing Geospatial Data: Distributed-Cloud Model, Relational Database, And Indexed Flat File, Matthew A. Toups

LSU New Orleans Theses and Dissertations

Geographic Information Systems (GIS) and related applications of geospatial data were once a small software niche; today nearly all Internet and mobile users utilize some sort of mapping or location-aware software. This widespread use reaches beyond mere consumption of geodata; projects like OpenStreetMap (OSM) represent a new source of geodata production, sometimes dubbed “Volunteered Geographic Information.” The volume of geodata produced and the user demand for geodata will surely continue to grow, so the storage and query techniques for geospatial data must evolve accordingly.

This thesis compares three paradigms for systems that manage vector data. Over the past few decades …

Test Plan For Real-Time Modeling & Simulation Of Single Pole Switching Relays, Ram Mohan Sanaboyina

LSU New Orleans Theses and Dissertations

A real-time simulator (RTS) with digital and analog input/output modules is used to conduct hardware-in-the-loop simulations to evaluate performance of power system equipment such as protective relays by exposing the equipment to the simulated realistic operating conditions. This work investigates the use of RTS to test relays with single-pole-switching (SPS) feature. Single-pole switching can cause misoperations due to fault arc during reclosing of the breakers. Through this investigation, a test procedure appropriate for the testing SPS relays has been developed. The test procedure includes power system modeling for real time simulation, relay test setup, and test plan. HYPERSIM real-time simulator …

Unequal A Priori Probability Multiple Hypothesis Testing In Space Domain Awareness With The Space Surveillance Telescope, Tyler J. Hardy, Stephen C. Cain, Travis F. Blake

Faculty Publications

This paper investigates the ability to improve Space Domain Awareness (SDA) by increasing the number of detectable Resident Space Objects (RSOs) from space surveillance sensors. With matched filter based techniques, the expected impulse response, or Point Spread Function (PSF), is compared against the received data. In the situation where the images are spatially undersampled, the modeled PSF may not match the received data if the RSO does not fall in the center of the pixel. This aliasing can be accounted for with a Multiple Hypothesis Test (MHT). Previously, proposed MHTs have implemented a test with an equal a priori prior …

Seismic Design Of A Prestressed Concrete Bridge, Alperen Ozel

LSU New Orleans Theses and Dissertations

Latest advancements in software have become an indispensable tool in structural analysis. The main goal of this thesis is to examine dynamic responses of bridges in seismic activity with the help of such tool as CSi Bridge. Therefore, throughout this study, a prestressed concrete bridge model will be thoroughly designed. In the first section, the required materials and structural components will be introduced. The following section will cover calculations required for modeling. The actual modeling of the structure will be carried out in Section 3. In Section 4, the set of required analysis for seismic design such as modal, pushover …

Volatile Organic Compound (Voc) Emission During Cured-In-Place-Pipe (Cipp) Sewer Pipe Rehabilitation, Elena Bourbour Ajdari

LSU New Orleans Theses and Dissertations

The maintenance or replacement of deteriorated pipes and culverts is a constant and significant concern for municipalities and transportation agencies in the United States (Donaldson and Wallingford, 2010). Trenchless technologies and especially the Cured-in-place pipe (CIPP) method have become increasingly common ways to preserve infrastructures owing to their feasibility, cost-effectiveness, and fewer social impacts (Jung and Sinha, 2007). Therefore, there is a growing need to understand the direct and indirect effects of pipeline rehabilitation activities on the environment. Nearly all past CIPP studies have focused on its mechanical properties, and its environmental impacts are poorly investigated and documented (Allouche et …

Odor Monitoring At The New Orleans East Bank Wastewater Treatment Plant, Felicia R. Figinsky

LSU New Orleans Theses and Dissertations

Hydrogen Sulfide is a corrosive, odorous, and hazardous gas that can cause issues for wastewater collection systems and treatment plants. When evaluating hydrogen sulfide concentrations it is important to consider the source, odor causing mechanism, and its level of toxicity. In this study, continuous hydrogen sulfide monitoring was performed for 8 locations within the New Orleans East Bank Wastewater Treatment Plant for a span of 5 months. Hot spots within the plant are located and data analysis is performed based on daily and hourly averages of hydrogen sulfide concentrations. The data was configured to show peak hydrogen sulfide concentrations at …

A Numerical Study Of Compressible Lid Driven Cavity Flow With A Moving Boundary, Amer Hussain

LSU New Orleans Theses and Dissertations

A two-dimensional (2-D), mathematical model is adopted to investigate the development of circulation patterns for compressible, laminar, and shear driven flow inside a rectangular cavity. The bottom of the cavity is free to move at a specified speed and the aspect ratio of the cavity is changed from 1.0 to 1.5. The vertical sides and the bottom of the cavity are assumed insulated. The cavity is filled with a compressible fluid with Prandtl number, Pr =1. The governing equations are solved numerically using the commercial Computational Fluid Dynamics (CFD) package ANSYS FLUENT 2015 and compared with the results for the …

Simulation And Performance Evaluation Of Algorithms For Unmanned Aircraft Conflict Detection And Resolution, Jeffrey H. Ledet

LSU New Orleans Theses and Dissertations

The problem of aircraft conflict detection and resolution (CDR) in uncertainty is addressed in this thesis. The main goal in CDR is to provide safety for the aircraft while minimizing their fuel consumption and flight delays. In reality, a high degree of uncertainty can exist in certain aircraft-aircraft encounters especially in cases where aircraft do not have the capabilities to communicate with each other. Through the use of a probabilistic approach and a multiple model (MM) trajectory information processing framework, this uncertainty can be effectively handled. For conflict detection, a randomized Monte Carlo (MC) algorithm is used to accurately detect …

Focal Adhesion Kinase Regulation In Stem Cell Alignment And Spreading On Nanofibers, Mohammad Nahid Andalib, Jeong Soon Lee, Ligyeom Ha, Yuris A. Dzenis, Jung Yul Lim

Department of Mechanical and Materials Engineering: Faculty Publications

While electrospun nanofibers have demonstrated the potential for novel tissue engineering scaffolds, very little is known about the molecular mechanism of how cells sense and adapt to nanofibers. Here, we revealed the role of focal adhesion kinase (FAK), one of the key molecular sensors in the focal adhesion complex, in regulating mesenchymal stem cell (MSC) shaping on nanofibers. We produced uniaxially aligned and randomly distributed nanofibers from poly(L-lactic acid) to have the same diameters (about 130 nm) and evaluated MSC behavior on these nanofibers comparing with that on flat PLLA control. C3H10T1/2 murine MSCs exhibited upregulations in FAK expression and …

Mesenchymal Stem Cell Fate: Applying Biomaterials For Control Of Stem Cell Behavior, Hilary J. Anderson, Jugal Kishore Sahoo, Rein V. Ulijn, Matthew J. Dalby

Publications and Research

The materials pipeline for biomaterials and tissue engineering applications is under continuous development. Specifically, there is great interest in the use of designed materials in the stem cell arena as materials can be used to manipulate the cells providing control of behavior. This is important as the ability to “engineer” complexity and subsequent in vitro growth of tissues and organs is a key objective for tissue engineers. This review will describe the nature of the materials strategies, both static and dynamic, and their influence specifically on mesenchymal stem cell fate.

3-D Shape Recovery From A Single Camera Image, Vijai Jayadevan, Aaron Michaux, Edward Delp, Zygmunt Pizlo

MODVIS Workshop

3-D shape recovery is an ill-posed inverse problem which must be solved by using a priori constraints. We use symmetry and planarity constraints to recover 3-D shapes from a single image. Once we assume that the object to be reconstructed is symmetric, all that is left to do is to estimate the plane of symmetry and establish the symmetry correspondence between the various parts of the object. The edge map of the image of an object serves as a good representation of its 2-D shape and establishing symmetry correspondence means identifying pairs of symmetric curves in the edge map. The …

Figure-Ground Organization Using 3d Symmetry, Aaron Michaux, Vijai Jayadevan, Edward Delp, Zygmunt Pizlo

MODVIS Workshop

We present a novel approach to object localization using mirror symmetry as a general purpose and biologically motivated prior. 3D symmetry leads to good segmentation because (i) almost all objects exhibit symmetry, and (ii) configurations of objects are not likely to be symmetric unless they share some additional relationship. Furthermore, psychophysical evidence suggests that the human vision system makes use symmetry in constructing 3D percepts, indicating that symmetry may be important in object localization. No general purpose approach is known for solving 3D symmetry correspondence in 2D camera images, because few invariants exist. Therefore, to test symmetry as a clustering …

Assessment Of Soil Washing For Simultaneous Removal Of Heavy Metals And Low-Level Petroleum Hydrocarbons Using Various Washing Solutions, Deok-Hyun Moon, Jae-Woo Park, Agamemnon Koutsospyros, Kyung Hoon Cheong, Yoon-Young Chang, Ki-Tae Baek, Raehyun Jo, Jeong-Hun Park

Civil Engineering Faculty Publications

Bench-scale soil washing experiments were conducted for simultaneous removal of heavy metals (Pb, Cu, Zn) and low-level petroleum hydrocarbon contaminants from soils. Various washing solutions including hydrochloric acid (HCl), nitric acid (HNO₃), sulfuric acid (H₂SO₄), tartaric acid (C₄H₆O₆) and ethylenediaminetetraacetic acid (C₁₀H₁₆N₂O₈, EDTA) were used. The concentration of the washing solutions ranged from 0.1 to 3M with a liquid-to-solid ratio of 10. The soil washing results showed that hydrochloric acid (HCl) was the best washing solution at 3M for heavy …

The Use Of Semi-Structured Interviews For The Characterisation Of Farmer Irrigation Practices, Jimmy O’Keeffe, Wouter Buytaert, Ana Mijic, N. Brozovic, Rajiv Sinha

Daugherty Water for Food Global Institute: Faculty Publications

For the development of sustainable and realistic water security, generating information on the behaviours, characteristics, and drivers of users, as well as on the resource itself, is essential. In this paper we present a methodology for collecting qualitative and quantitative data on water use practices through semi-structured interviews. This approach facilitates the collection of detailed information on actors’ decisions in a convenient and cost-effective manner. Semi-structured interviews are organised around a topic guide, which helps lead the conversation in a standardised way while allowing sufficient opportunity for relevant issues to emerge. In addition, they can be used to obtain certain …