Biochemical and Biomolecular Engineering Commons^™

Investigating The Detection And Transport Of Low Molecular Weight Species To Better Understand Their Impact On Disease Pathology And Treatment, Haley Michelle Duncan

Graduate Theses and Dissertations

Due to the drastic differences in the projects involved, this dissertation will be split into two sections: Alzheimer’s disease and chronic kidney disease. Each chapter will contain an introduction, materials and methods section, results and discussion, conclusions, and works cited section. The Alzheimer’s disease chapter will also have a supplemental figures section containing the polyacrylamide gels that were silver stained and quantified via ImageJ. The presence of elevated levels of amyloid beta oligomeric species in the brain has been related to the pathology of Alzheimer’s disease. As these oligomeric species are transient in nature, their kinetics pose a challenge to …

Yeast Surface Display Of Multiple Heterologous Proteins Via Molecular And Biochemical Processing Techniques, Shadrach Ibinola

Graduate Theses and Dissertations

The arming of yeast cell wall, also known as yeast surface display, is an effective technology for expressing target proteins extracellularly. It has been shown to provide bioengineered yeast with qualities that are not known to native yeast such as the ability to survive harsh conditions including high pH and temperature, for example. Furthermore, this technique has been implemented for various applications which include, protein engineering, bioethanol production, bioremediation, vaccine construction, and antibody development. The major mechanisms of yeast surface display exploit agglutination, flocculation, or proteins with internal repeats as the means by which a target (protein) is covalently attached …

Developing Anisotropic Soft Matter As Functional Materials To Interface With Biological Systems, Homa Ghaiedi

Graduate Theses and Dissertations

Liquid crystals (LCs) constitute a subset of soft matter possessing hybrid characteristics from both liquid and crystalline phases. In this state, the center of masses remains uncorrelated similar to an isotropic phase, while the long axis of molecules aligns in parallel, resulting in molecules exhibiting orientational and positional order, either individually or concurrently. The directional alignment inherent in LCs arises from the asymmetrical shape and structure of the constituent molecules, causing a spontaneous self-assembly into ordered phases. This cooperative molecular behavior manifests distinctive optical features when subjected to cross polarizers. The remarkable sensitivity of LCs to external stimuli, including temperature, …

Dpd Guided Insight On The Formation Process Of Polyethersulfone Membranes By Nonsolvent Induced Phase Separation And The Effects Of Additives, Eric Ledieu

Graduate Theses and Dissertations

Dissipative particle dynamics (DPD), a coarse grain simulation method, was applied to the membrane formation process of non-solvent induced phase separation (NIPS) to gain further insight on the mechanism of certain variables and how they affect the final morphology. NIPS involves two solutions, an organic polymer dissolved in an organic solvent colloquially called the dope and an aqueous coagulation bath, brought into contact with one another. The solvents then mix, causing the polymer to fall out of solution as an asymmetric membrane with a dense surface layer and a more open subsurface layer in response to the decreasing solubility. Polyethersulfone …

Polymeric Biomaterials Approaches For Engineering The In Vitro Cellular Microenvironment For Mscs, Mahsa Letter-Mahsa Haseli

Graduate Theses and Dissertations

Cell therapy is a technology that relies on replacing diseased or dysfunctional cells with healthy functioning ones. One of the cells used for such advanced therapies are stem cells, owing to their ability to differentiate into specific cells required for repairing damaged or defective tissues or cells. The majority of cell-based products are intended to transiently persist in the patient, secreting factors which then allow the patient’s body to heal; in these products, the cells are subsequently eliminated from the body. Furthermore, unique manufacturing platforms, in addition to novel commercialization strategies, will be required to create a successful, sustainable cell …

Enhancing Human Schwann Cells Reparative Behavior Using Heparin/Collagen Layer-By-Layer Coatings, Luis Carlos Pinzon-Herrera

Graduate Theses and Dissertations

When a peripheral nerve injury (PNI) occurs, the gold standard for tissue regeneration is the use of autografts. However, due to the secondary effects produced by multiple surgeries involved in the removal and implantation of autografts for very small lesions, it is possible to replace them with the use of Nerve Guide Conduits (NGCs). However, NGCs are limited to short lesions (less than 1 cm). This limitation is caused by the absence of compounds in the extracellular matrix (ECM) that autografts can provide. Since much of the regenerative process takes place on the NGC surface, our work aims to modify …

Life Cycle Assessment Of Novel Electrochemical Struvite Recovery In Municipal Wastewater Treatment At The Plant And U.S. Watershed Scales, Karla Morrissey

Graduate Theses and Dissertations

As the supply of phosphorus decreases, nutrient recovery in municipal wastewater treatment has become an important area of research. Recovery methods for phosphorus-containing fertilizers, such as struvite, and other products are currently being developed and assessed for their efficiency and economic feasibility. Potential environmental impacts from nutrient recovery are also being assessed as design for the environment is important, especially with regards to the eutrophying potential of phosphorus in phosphorus-limited freshwater aquatic systems. Life cycle assessment (LCA) is a tool that can be used to estimate potential environmental impacts of a product or service focusing on its entire life cycle. …

Greenhouse Tomatoes: Process Simulation, Juan Gabriel Marin Jr.

Graduate Theses and Dissertations

Growing population demand and challenges brought on by climate change have spurred the need for more resilient fruit and vegetable supply chains. One agricultural technology of significant interest is the use of greenhouses for food production. Greenhouses create a stable and adaptable environment for crops such as tomatoes to grow year-round. Fresh tomatoes are the second most consumed vegetable per capita in U.S. diets, currently averaging 20.7 pounds. The growing consumption of fresh tomatoes has been the result of increasing cultural diversity in the United States.

To meet the growing demand, Venlo-type greenhouses have been frequently used by growers. It …

Recombinant Production And Purification Of Green Fluorescent Protein (Gfp)-Fused Metal Binding Protein For Palladium Nanoparticle Synthesis, Shadrach Ibinola

Graduate Theses and Dissertations

In lieu of chemical and physical methods, biologically guided synthesis is increasingly used as a cost-effective medium for the fabrication of nanoparticles (NP). Recently, a palladium metal binding sequence Pd4 (TSNAVHPTLRHL) has been demonstrated to be instrumental in the production of palladium (Pd) nanoparticles. Although, by eliminating the additional cost of purification of the protein, the crude lysate of E. coli containing Pd specific protein has been proven to be a viable cost-effective means for the synthesis of Pd NP, studies have not be done to ascertain the comparative catalytic activity of nanoparticles synthesized from both clarified lysate and pure …

Investigation Of Membrane Based Processes For Biomedical Applications, Efecan Pakkaner

Graduate Theses and Dissertations

As substantial developments were achieved in nanotechnology and polymer engineering, especially in the last few decades, the use of membranes and membrane-based procedures was found to be expanding into more and more research and development areas; including biological engineering, life sciences and biomedical engineering. Not only have they been the main focus of meaningful research, but they have also been the main pieces of the solutions to very thorny problems encountered within a wide range of applications from microfluidics to water treatment, thanks to their versatility, cost-effectiveness and biocompatibility, when compared to conventional separation techniques. To celebrate and embrace these …

A Portable Point-Of-Care Device Using Joule Heating And Latent Energy Storage For The Temperature Regulation Of Isothermal Nucleic Acid Amplification Tests, Aubrey Lynn Schultz

Graduate Theses and Dissertations

Accurate and early diagnosis of infectious diseases extremely important. Rapid diagnosis allows for effective treatment and increases the chance for recovery without complications. Additionally, the ability to test the populace frequently, swiftly, and affordably significantly aids in containing wide-scale outbreaks. In terms of specificity and sensitivity, nucleic acid amplification tests (NAAT) are one of the best options for diagnosing infectious diseases. Isothermal NAATS present a unique opportunity to create diagnostic tests deployed at a Point-of-Care (POC) level. Specifically, loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) have the potential to deliver reliable POC diagnostics in low-resource settings. When designing …

Investigative Study Of Microalgal And Electrochemical Wastewater Treatment Systems And Modeling Of The Wafer-Enhanced Electrodeionization Using Supervised Learning, Humeyra Betul Ulusoy Erol

Graduate Theses and Dissertations

Wastewater has a serious impact on environment and public health due to its high concentration of nutrients and toxic contaminants. Without proper treatment, excess nutrients discharged in wastewater can cause a damage to the ecosystem such as undesirable pH shifts, cyanotoxin production, and low dissolved oxygen concentrations.

Main objectives of this dissertation work were to investigate i) the biofuel potential of P. cruentum when grown in swine wastewater, ii) the influence of four most commonly used ion exchange resins on the system efficiency and selectivity for the removal of sodium, calcium, and magnesium ions, and iii) the modeling of wafer-enhanced …

Peptoid And Antibody-Based Gfp Sensors, Solomon Isu

Graduate Theses and Dissertations

In this work, we have made and characterized a pair of immunobiosensors for detecting the green fluorescent protein (GFP) in an aqueous matrix. An anti-GFP antibody-based biosensor was assembled to detect GFP, while a novel peptoid (N-substituted oligomers of glycine designated as IOS-1) biosensor was also assembled for GFP detection. A quartz crystal microbalance (QCM) gold sensor was used as the supporting substrate for self-assembly of the immunobiosensors. Gravimetric measurements of the QCM gold sensor during immunobiosensor construction and operation were available in real-time using a QCM instrument. X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and Fluorescence microscopy were used …

Development Of A Comsol Microdialysis Model, Towards Creation Of Microdialysis On A Chip With Improved Geometries And Recovery, Patrick Pysz

Graduate Theses and Dissertations

Microdialysis (µD) sampling is a diffusion-limited sampling method that has been widely used in different biomedical fields for greater than 35 years. Device calibration for in vivo studies is difficult for current non-steady state analytes of interest correlated with both inflammatory response and microbial signaling molecules (QS); which exist in low ng/mL to pg/mL with molecular weights over a wide range of 170 Da to 70 kDa. The primary performance metric, relative recovery (RR), relating the collected sample to the extracellular space concentration varies from 10% to 60% per analyte even under controlled bench-top conditions. Innovations in microdialysis device design …

Bioengineering Extracellular Matrix Scaffolds For Volumetric Muscle Loss, Kevin Roberts

Graduate Theses and Dissertations

Volumetric muscle loss overwhelms skeletal muscle’s ordinarily capable regenerative machinery, resulting in fibrosis and severe functional deficits which have defied clinical repair strategies. My work spans the design and preclinical evaluation of implants intended to drive the cell community of injured muscle toward a regenerative state, as well as the development of an understanding of the molecular responses of this cell community to biomaterial interventions. I demonstrate a new class of biomaterial by leveraging the productive capacity of sacrificial hollow fiber membrane cell culture; I show specifically that unique threads of whole extracellular matrix can be isolated by solvent degradation …

Self-Assembled Nanoantenna Enhance Optical Activity And Transport In Scalable Thin Films And Interfaces, Keith Richard Berry Jr.

Graduate Theses and Dissertations

Continued population growth and the decrease of existing energy platforms demands long-term solutions for development and implementation of scalable plasmonic metamaterials for energy and agricultural applications. Self-assembled nanoantenna into random and ordered arrangements are advanced herein for optical and thermal enhancements in scalable thin film. An analytical approach to estimating the thermal dynamics of random arrangements of nanoantenna resulted in estimates within 30% across a range of geometric parameters, nanoantenna-containing media, and thermal parameters. Multimodal thermal dynamics of polymer thin films containing gold nanoparticles (AuNPs) were observed through the natural log of the dimensionless temperature driving force plotted versus time …

Peptoid-Modified Bicelles As Surrogate Cell Membranes For Membrane Protein Sensors And Analytics, Helya Najafi

Graduate Theses and Dissertations

Membrane-affiliated interactions are significant in understanding cell function, detecting biomarkers to diagnose disease, and in testing the efficiency of new therapeutic targets. Model membrane systems have been developed to study membrane proteins, allowing for stable protein structure and maintaining native activity. Bicelles, disc-shaped lipid bilayers created by combining long- and short-chain phospholipids, are the model membrane system of focus in this study. Bicelles are accessible from both sides and have a wide size range, which make them attractive for studying membrane proteins without affecting function. In this work, bicelles were functionalized with two peptoids to alter the edge and face …

Effects Of Filtration Conditions On Clearance Of Bacteriophage, Rigen Te

Graduate Theses and Dissertations

Virus Clearance is one of the major challenges in biopharmaceutical industry, especially in the manufacturing of drugs. U.S. Food and Drug Administration (FDA) recommends using several model bacteriophages as surrogates of pathogenic mammalian virus for process validation. In this work, two bacteriophages were tested against 30 kDa, 100 kDa, 300 kDa polyethersulfone (PES) membranes to investigate the effects of pore size on virus clearance. Virus particles were spiked into the protein feed solutions containing bovine serum albumin (BSA) or lysozyme at different concentrations. Besides protein concentration, the effects of feed pH on the filtration performance and virus rejection were also …

Peptoid-Based Microsphere Coatings For Use As Tunable Biocompatible Interfaces, German Raul Perez Bakovic

Graduate Theses and Dissertations

The pursuit of sensitive, non-invasive, and cost efficient diagnostic tools for early stage disease detection have led to the development of sophisticated biosensor technologies for proteomic studies. As these markers increase in complexity, the role of support substrates grows increasingly important. Limitations in existing support substrates include the potential for increased sensitivity, binding specificity, and bio-stability. Ideal support substrates need to provide biocompatible and bioresistant surfaces, that offer high surface areas for binding, and enables the incorporation of diverse chemistries. The use of peptoids as the basis for the deposition of uniform microsphere coatings offers a mean to the attainment …

Membrane Chromatography For Bioseparations: Ligand Design And Optimization, Zizhao Liu

Graduate Theses and Dissertations

Membrane chromatography, or membrane adsorber, represents an attractive alternative to conventional packed bed chromatography used in downstream processing. Membrane chromatography has many advantages, including high productivity, low buffer consumption and ease to scale up. This doctoral dissertation focuses on developing novel polymeric ligands for protein separations using membrane chromatography. Atom transfer radical polymerization (ATRP), known as a controlled radical polymerization technique, has been used to control the architecture of grafted polymeric ligands. The center theme of this dissertation is to develop new polymeric ligands and investigate how the polymer’s property (e.g. flexibility, hydrophobicity) and architecture (e.g. chain density, chain length) …

Cloning, Fed-Batch Expression And Purification Of A Novel Anti-Candida Peptide And Development Of A Cleavage Resistant Variant Of Green Fluorescence Protein, Rudra Palash Mukherjee

Graduate Theses and Dissertations

This work illustrates that the intelligent design of a bioprocess for the production of peptides of various length is possible by having a prior knowledge of desired product, cell line, fermentation conditions, and choices for downstream recovery. Cloning, expression, and recovery of a novel 12-mer anti-Candida peptide served as an illustrative case for a proposed Escherichia coli platform. The antifungal peptide, expressed as a fusion to GFPUV during high call density fed-batch, was recovered from cell lysate, concentrated using ion exchange chromatography, and finally cleaved with cyanogen bromide. The platform was capable of producing active peptide capable of arresting the …

Effects Of Magnetically Induced Micro-Mixing On Nanofiltration Performance, Guanghui Song

Graduate Theses and Dissertations

Nanofiltration (NF) is a relatively new membrane separation process mainly used for removing low molecular weight species from aqueous and non-aqueous solutions. NF membranes suffer from concentration polarization leading to membrane fouling thus compromised membrane performance. Magnetically responsive nanofiltration (NF) membranes functionalized with superparamagnetic nanoparticles (SPNs) attached to the chain ends of grafted polymer nanolayers have been shown to be effective in breaking concentration polarization at the membrane-liquid interface under an appropriate external oscillating magnetic field. Under an oscillating magnetic field, the movement of the polymer chains acts as micro-mixer leading to the suppression of concentration polarization and improved filtration …

Using Peptoids To Build Robust, Efficient Microarray Systems, Dhaval Sunil Shah

Graduate Theses and Dissertations

Recent studies have shown microarrays to be indispensable for various biological applications, allowing for high-throughput processing and screening of biological samples such as RNA, DNA, proteins and peptides using a small sample volume (< 1 µL). Peptoids (poly-N-substituted glycine oligomers) can be used as a substitute for antibodies as capture molecules, as well as coatings for slides in antibody microarrays. The ease of synthesis of peptoids, high customizability with desired bioactivity, and speed of synthesis allows us to build a diagnostic system with a large dynamic range that can detect biomolecules from a minimal sample size. In this study, peptoid-based antibody mimics are designed to have both structural and functional features similar to those of antibodies, including a stable constant region (scaffolding) and a variable region for protein recognition. Peptoids previously screened via combinatorial library synthesis to be specific to bind Mdm-2 (mouse double minute 2 homolog) and GST (gluthathione S-transferase), have been synthesized. The protein recognition peptoids have been conjugated to PEG (polyethylene glycol) molecules with modified end groups; an amine group on one end that allows for immobilization and orientation on the slide, and an azide group on the other end that will allow for attachment to the peptoid through “click chemistry”. The number of capture molecules printed on the slides can be increased by making the available surface area of the slide larger via coating with microspheres. We have determined that partially water soluble peptoids that are also helical, can self-assemble into microspheres. Sequences have been developed that can consistently produce uniform microsphere coatings on slides that increase the overall surface area. A high surface area corresponds to a higher number of binding sites, and therefore a more sensitive system. The work done has shown that slides may be successfully coated in order to potentially improve the detection system.

Peptoid-Based Klvff Mimics: A Unique Approach To Alzheimer's Disease, James Phillip Turner

Graduate Theses and Dissertations

Alzheimer’s disease (AD) is the leading form of dementia worldwide. AD patients experience a slow, gradual cognitive decline that includes loss of memory and behavioral stability as the disease progresses. Surprisingly, AD is the sixth leading cause of death in the United States and has had a profound impact on the U.S. economy. Though there are medications to help improve the quality of life of diagnosed patients for a period of time, there is still no cure for AD. AD is characterized by the build-up of amyloid plaques that develop from the aggregation of the amyloid beta protein (Aβ) in …

The Production Of Biobutanol From Biomass Via A Hybrid Biological/Chemical Process, Thomas Melvin Potts

Graduate Theses and Dissertations

Biobutanol use as a fuel began in the late 19th century. Problems remain in economic viability. A review of the state of the art and need for technical advances is presented.

The technical potential of producing biofuel from a naturally occurring macroalgae was studied. The algae grow in Jamaica Bay, New York City, in contaminated water. The process consisted of mechanical harvesting, drying, grinding, and acid hydrolysis to form an algal sugar solution. Clostridium beijerinckii and C. saccharoperbutylacetonicum were used in an acetone butanol ethanol (ABE) fermentation to make butanol. Fermentation was followed by distillation Butanol concentrations during fermentation reached …

Application Of Membrane Processes For Concentration And Separation Of Sugar Streams In Biofuel Production, Mohammadmahdi Malmali

Graduate Theses and Dissertations

The overall objective of this study was identification and development of a sugar concentration/separation membrane filtration unit to improve the bioconversion of lignocellulosic biomass into chemicals and fuels. This thesis is divided into three main parts. The first part is about our studies on the use of nanofiltration membranes for concentration of sugars in a lignocellulosic biomass hydrolysate. In addition, the feasibility of simultaneous removal of acetic acid, 5-(hydroxymethyl)furfural and furfural from the hydrolysate has also been investigated. The results obtained indicate that both concentration of sugars and removal of hydrolysis degradation products is feasible. However, careful selection of the …

Comparing Virus Ultrafiltration Of Bacteriophages With Filtration Of Minute Virus Of Mice, Kent Smith

Graduate Theses and Dissertations

Industrial production of protein therapeutics demand rigorous testing and clearance of viruses. The U.S. Food and Drug Administration dictate the purity of pharmaceuticals with regards to viral contamination. As this testing is time consuming and expensive using mammalian cells and viruses, bacteriophages may provide a faster and cheaper alternative for membrane filtration processes. We used ultrafiltration membranes to filter protein solutions with viruses. Two bacteriophages were tested against membranes with two different pore sizes. These membranes were then tested by inverting the membrane's orientation. Flux measurements and log virus removal data were taken. Flux and log virus removal were seen …

An Experimental And Computational Evaluation Of The Importance Of Molecular Diffusion In Gas Gravity Currents, Jeremy Jac Herman

Graduate Theses and Dissertations

The accidental release of hazardous, denser-than-air gases during their transport or manufacture is a vital area of study for process safety researchers. This project examines the importance of molecular diffusion on the developing concentration field of a gas gravity current released into a calm environment. Questions which arose from the unexpectedly severe explosion in 2005 at Buncefield, England were of particular interest. The accidental overfilling of a large tank with gasoline on a completely calm morning led to a massive open air explosion. Forensic evidence showed that at the time of ignition, a vapor cloud, most of which now appears …

Characterization And Quantification Of Monomers, Oligomers, And By-Products From Switchgrass Hemicelluloses During Biomass Pretreatment, Kris Allan Bunnell

Graduate Theses and Dissertations

Production of fuels and chemicals from biomass is contingent upon economical release of carbohydrates from biomass. Carbohydrates can then be used for production of bio-based products using a biochemical conversion process. Pretreatment, the first step of the biochemical conversion process, has been suggested to be the most costly step of the conversion process. Thus, better understanding the behavior of biomass during pretreatment is imperative for an economically viable production of biofuels and chemicals. Elucidating the physicochemical properties of biomass and developing an understanding the depolymerization patterns of biomass during pretreatment will help progress towards this goal.

In this study, July- …

Algae To Propellants: Biosynthesis Of 1,2-Propanediol From Algal Derived Sugars, Lauren Rachelle Merriman

Graduate Theses and Dissertations

Propylene glycol dinitrate (PGDN) is an important component used in the formulation of energetic materials. A renewable, environmentally-friendly and effective production technique for the production of 1,2-propanediol, an important precursor to PGDN, is desired. While fermentation of 1,2-propanediol has been achieved, it suffers from major limitations such as expensive feed sugars and relatively low yields. It is hypothesized that algae sugars could serve as a feedstock for the microbial synthesis of 1,2-propanediol, thereby reducing production costs and enhancing sustainability.

Algal biomass produced from an Algal Turf Scrubber® has proven to have significant potential as a source of fermentable materials. However, …