Biomedical Engineering and Bioengineering Commons^™

Continuous Production Of Proteins: Integration Of Polishing Using Mcsgp, Fabian Steinebach, Daniel Karst, Massimo Morbidelli

Integrated Continuous Biomanufacturing II

Continuous counter-current processes for polishing steps overcome the purity-yield tradeoff of classical batch chromatography by internal recycling of overlapping regions of the chromatogram. Particularly in the case of challenging separation tasks this leads for the same product quality to significantly higher yields and productivity. Processes using these fundamental principals have been successfully implemented in the chemical industry up to multi-ton production.

In an integrated continuous process, with a perfusion bioreactor and a continuous capture process, the product quality of the capture product is different compared to classical batch processing. The lower residence time in the bioreactor and the higher loading …

Exometabolome Characterization Of High Cell Density Culture Perfusion And Optimization Of The Cell Specific Perfusion Rate, Veronique Chotteau, Leila Zamani, Ye Zhang, Magnus Aberg, Anna Lindahl, Axel Mie

Integrated Continuous Biomanufacturing II

High cell density perfusion has the potential to generate intensified processes. A target for such perfusion process can be to maintain a steady state culture around 100 x 10⁶ cells/mL. A challenge for the development of these processes is to minimize the medium consumption, to reduce the costs associated with this raw material and with the harvest processing. This is particularly true for processes at densities such as 100 x 10⁶ cells/mL for which several reactor volumes per day are typically necessary.

We have shown that cell specific perfusion rate (CSPR) is an approach, which has proven to …

Continuous Downstream Processing Of A Monoclonal Antibody Using Periodic Counter Current Chromatography (Pcc) And Straight Through Processing (Stp), Hans Blom, Christer Eriksson, Annika Forss, Helena Skoglar

Integrated Continuous Biomanufacturing II

There is an increased interest to perform process intensification in order to reduce costs and improve throughput in the development and production of monoclonal antibodies (MAbs). One solution to these demands can be to implement continuous or semi-continuous downstream processing. New emerging technologies such as periodic counter-current (PCC) chromatography and straight through processing (STP) are entering the market. Here, these two technologies were evaluated in a continuous three step chromatography MAb process. The capture step was performed with protein A media (resin) on a 3 column PCC chromatography system followed by two polishing steps which were connected in series with …

Integrated Solutions For Continuous Processing In Mobius® Bioreactor Systems, Andrew Clutterbuck

Integrated Continuous Biomanufacturing II

Throughout the bioprocess industry, the use of disposable systems such as bioreactors is playing an increasingly significant role. Manufactures are recognising the strategic benefits of single use systems in both reducing production costs and increasing overall manufacturing throughput.

Over the past few years there has been increasing demand for larger scale disposable bioreactor systems, to satisfy the manufacturing demands and an emphasis on suppliers such as Merck Millipore to demonstrate robust and reproducible scale-up within these systems.

This presentation will explore the design philosophies behind the Mobius^® bioreactors and will also give an overview of the novel SensorReady external …

Optical Enzymatic Sensors For Continuous Monitoring Of Bioreactors, Kenneth Reardon, Brian Heinze, Trenton Danna

Integrated Continuous Biomanufacturing II

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. An overview of sensors will be presented, focusing on in-situ sensors that contact the biological medium, and external sensors that contact the medium either optically (ex situ) or via a sterile (and disposable) …

Salt-Tolerant Cation Exchange Hd-Sb Hydrogel Membrane: Mab Purification Performance In Flowthrough Mode, Dharmesh Kanani, Daniel Luo, Nirali Paghdal, Navneet Sidhu, Annabel Shang, Jim Stout

Integrated Continuous Biomanufacturing II

Development of Protein A based purification platforms have simplified the downstream processing of monoclonal antibodies (mAb), the largest component of biopharmaceuticals. The ever increasing titer of the cell culture process is putting more pressure on the downstream process to further increase its productivity. The classical Protein A based mAb purification platform consists of two polishing steps, bind and elute cation-exchanger and flowthrough anion-exchanger. Cation-exchange chromatography is very efficient at the separation of HCP, leached Protein A and product-related impurities such as aggregates and fragments in bind and elute mode. However, anion-exchange chromatography is a proven technology to remove DNA, viruses, …

Glycoexpress: A Toolbox For The High Yield Production Of Glycooptimized Fully Human Biopharmaceuticals In Perfusion Bioreactors At Different Scales, Steffen Kreye

Integrated Continuous Biomanufacturing II

Background and Novelty: Glycosylation is one of the major post-translational modifications of biotherapeutics important for bioactivity, bioavailability, immunogenicity and patient coverage. By establishment of the GlycoExpress™ toolbox (GEX™) we have generated a set of glycoengineered human cell lines for the high yield production of fully human glycoproteins to optimize the glycosylation of antibodies and non-antibody biotherapeutics for improvement of the clinical efficacy and side effects. The system is biotechnologically superior in quality, reproducibility and yield compared to other including conventional production systems. All four clinical products derived from GlycoExpress cells are produced using a perfusion bioreactor system in order to …

Designing A Microbial Cultivation Platform For Continuous Biopharmaceutical Production, Nicholas Mozdzierz, Kerry Love, Alan Stockdale, John Clark, Noelle Colant, Christopher Love

Integrated Continuous Biomanufacturing II

The existing biopharmaceutical manufacturing paradigm is poorly suited to produce biologic drugs on demand at a point-of-care. Generally, commercial-scale (~2,000 - 10,000 L) manufacturing using fed-batch cultivation and fixed stainless-steel infrastructure is concentrated in developed nations and results in process cycle times on the order of weeks to months.^1,2 Coupled with the complex logistical challenges associated with continuous “plant-to-patient” cold-chains, the geographically biased nature of therapeutic protein production today can limit access to biologic drugs in developing areas of the world.³ There is an opportunity to create technologies capable of rapidly generating biopharmaceuticals in situ in emergency situations, …

Robust Design And Operation Of Quasi-Continuous Adenovirus Purification By Two-Column, Simulated Moving-Bed, Size-Exclusion Chromatography, Jose Mota, Ricardo Silva, Piergiuseppe Nestolla, Cristina Peixoto, Manuel Carrondo

Integrated Continuous Biomanufacturing II

Adenoviruses (Ads) are considered one of the most suitable platforms for production of viral vaccines and gene therapy vectors. Their broad tissue tropism and large transgene packing capacity make them attractive candidates for innovative virotherapies. Ads can be produced in a complementary cell line in both adherent and suspension culture systems, such as HEK-293 or PER-C6 cells, or A549 for oncolytic therapies.

Liquid chromatography is currently the core technique for vector purification, and its use is often integrated vertically within the downstream processing (DSP) strategy, as it easily fits into the early capture stage as well as into the final …

Final Program, Kathy Chan

Integrated Continuous Biomanufacturing II

No abstract provided.

Gaussian Nonlinear Line Attractor For Learning Multidimensional Data, Theus H. Aspiras, Vijayan K. Asari, Wesam Sakla

Electrical and Computer Engineering Faculty Publications

The human brain’s ability to extract information from multidimensional data modeled by the Nonlinear Line Attractor (NLA), where nodes are connected by polynomial weight sets. Neuron connections in this architecture assumes complete connectivity with all other neurons, thus creating a huge web of connections. We envision that each neuron should be connected to a group of surrounding neurons with weighted connection strengths that reduces with proximity to the neuron. To develop the weighted NLA architecture, we use a Gaussian weighting strategy to model the proximity, which will also reduce the computation times significantly.

Once all data has been trained in …

Computational Characterization Of Exogenous Micrornas That Can Be Transferred Into Human Circulation, Jiang Shu, Kevin Chiang, Janos Zempleni, Juan Cui

School of Computing: Faculty Publications

MicroRNAs have been long considered synthesized endogenously until very recent discoveries showing that human can absorb dietary microRNAs from animal and plant origins while the mechanism remains unknown. Compelling evidences of microRNAs from rice, milk, and honeysuckle transported to human blood and tissues have created a high volume of interests in the fundamental questions that which and how exogenous microRNAs can be transferred into human circulation and possibly exert functions in humans. Here we present an integrated genomics and computational analysis to study the potential deciding features of transportable microRNAs. Specifically, we analyzed all publicly available microRNAs, a total of …

An Immersed Boundary Geometric Preprocessor For Arbitrarily Complex Terrain And Geometry, Inanc Senocak, Micah Sandusky, Rey Deleon, Derek Wade, Kyle Felzien, Marianna Budnikova

Mechanical and Biomedical Engineering Faculty Publications and Presentations

There is a growing interest to apply the immersed boundary method to compute wind fields over arbitrarily complex terrain. The computer implementation of an immersed boundary module into an existing flow solver can be accomplished with minor modifications to the rest of the computer program. However, a versatile preprocessor is needed at the first place to extract the essential geometric information pertinent to the immersion of an arbitrarily complex terrain inside a 3D Cartesian mesh. Errors in the geometric information can negatively impact the correct implementation of the immersed boundary method as part of the solution algorithm. Additionally, the distance …

Next-Generation Mrna Sequencing Reveals Pyroptosis-Induced Cd4+ T Cell Death In Early Simian Immunodeficiency Virus-Infected Lymphoid Tissues, Wuxun Lu, Andrew J. Demers, Fangrui Ma, Guobin Kang, Zhe Yuan, Yanmin Wan, Yue Li, Jiangqing Xu, Mark Lewis, Qingsheng Li

Nebraska Center for Biotechnology: Faculty and Staff Publications

Lymphoid tissues (LTs) are the principal sites where human immunodeficiency virus type 1 (HIV-1) replicates and virus-host interactions take place, resulting in immunopathology in the form of inflammation, immune activation, and CD4+ T cell death. The HIV-1 pathogenesis in LTs has been extensively studied; however, our understanding of the virus-host interactions in the very early stages of infection remains incomplete. We investigated virus-host interactions in the rectal draining lymph nodes (dLNs) of rhesus macaques at different times after intrarectal inoculation (days postinoculation [dpi]) with simian immunodeficiency virus (SIV). At 3 dpi, 103 differentially expressed genes (DEGs) were detected using next-generation …

Computational Replication Of The Patient-Specific Stenting Procedure For Coronary Artery Bifurcations: From Oct And Ct Imaging To Structural And Hemodynamics Analyses, Claudio Chiastra, Wei Wu, Benjamin Dickerhoff, Ali Aleiou, Gabriele Dubini, Hiromasa Otake, Francesco Migliavacca, John F. Ladisa Jr.

Biomedical Engineering Faculty Research and Publications

The optimal stenting technique for coronary artery bifurcations is still debated. With additional advances computational simulations can soon be used to compare stent designs or strategies based on verified structural and hemodynamics results in order to identify the optimal solution for each individual’s anatomy. In this study, patient-specific simulations of stent deployment were performed for 2 cases to replicate the complete procedure conducted by interventional cardiologists. Subsequent computational fluid dynamics (CFD) analyses were conducted to quantify hemodynamic quantities linked to restenosis.

Patient-specific pre-operative models of coronary bifurcations were reconstructed from CT angiography and optical coherence tomography (OCT). Plaque location and …

Biomechanical Investigation Of Elite Place-Kicking, Chase M. Pfeifer

Department of Mechanical and Materials Engineering: Dissertations, Theses, and Student Research

Many studies aim to understand the fundamentals of kicking commonly displayed by soccer players [4,6,10,16,17,18,24,25,28,29,30,34,36,38,40]. Of those studies, most are limited to a two-dimensional (2D) analysis using high-speed cameras for position tracking or utilizing electromyography to observe the activity of select muscles [4,6,18,25,29,36]. The few studies that investigate kicking using a three-dimensional (3D) model are limited in their position tracking capabilities and focus mainly on joint flexion potentials and foot speed.

This dissertation is a comprehensive biomechanical analysis (kinematic and EMG) of the field-goal place-kicking techniques of four elite kickers in American football. Data were compared and contrasted with ball …

Effect Of Imperceptible Vibratory Noise Applied To Wrist Skin On Fingertip Touch Evoked Potentials – An Eeg Study, Na Jin Seo, Kishor Lakshminarayanan, Leonardo Bonilha, Abigail W. Lauer, Brian D. Schmit

Biomedical Engineering Faculty Research and Publications

Random vibration applied to skin can change the sense of touch. Specifically, low amplitude white-noise vibration can improve fingertip touch perception. In fact, fingertip touch sensation can improve even when imperceptible random vibration is applied to other remote upper extremity areas such as wrist, dorsum of the hand, or forearm. As such, vibration can be used to manipulate sensory feedback and improve dexterity, particularly during neurological rehabilitation. Nonetheless, the neurological bases for remote vibration enhanced sensory feedback are yet poorly understood. This study examined how imperceptible random vibration applied to the wrist changes cortical activity for fingertip sensation. We measured …

Modeling Viscoelastic Behavior In Compact Bone Through A Distribution Of Collagen D-Spacing: A Finite Element Analysis, Christopher Ha

Master's Theses

Osteoporosis affects nearly 54 million people in the United States. The cost associated with treatment is estimated to be $19 billion per year and is expected to grow yearly. D-spacing is the staggering of collagen molecules found at the nanoscopic level. Previously thought to have a constant value, recent studies have found that D-spacing has a distribution of values throughout the tissue. As part of an ongoing effort in understanding the mechanisms that are affected by osteoporosis, a finite element model was developed to explore the effects of D-spacing distribution on the viscoelastic material properties of bone tissue. The goal …

Detection Of Acute Nervous System Injury With Advanced Diffusion-Weighted Mri: A Simulation And Sensitivity Analysis, Nathan P. Skinner, Shekar N. Kurpad, Brian D. Schmit, Matthew D. Budde

Biomedical Engineering Faculty Research and Publications

Diffusion-weighted imaging (DWI) is a powerful tool to investigate the microscopic structure of the central nervous system (CNS). Diffusion tensor imaging (DTI), a common model of the DWI signal, has a demonstrated sensitivity to detect microscopic changes as a result of injury or disease. However, DTI and other similar models have inherent limitations that reduce their specificity for certain pathological features, particularly in tissues with complex fiber arrangements. Methods such as double pulsed field gradient (dPFG) and q-vector magic angle spinning (qMAS) have been proposed to specifically probe the underlying microscopic anisotropy without interference from the macroscopic tissue organization. …

Smartphone Based 3d Printed Colorimeter For Biomedical Applications, Karthik Raj Konnaiyan

USF Tampa Graduate Theses and Dissertations

Here we present a novel Smartphone-based colorimeter and demonstrate its application to the measurements of glucose and protein concentrations in biological samples. The key innovation of our approach was to combine powerful image processing encoded into a mobile phone application with a low cost 3D printed sample holder that allowed to control lighting conditions and significantly improved sensitivity. Different solutions with protein and glucose concentrations ranging from 0 to 2000 mg/dL were prepared and analyzed using our system. The Smartphone-based colorimeter always correctly classified the corresponding reagent strip pads, what confirms that it can be used as a low cost …