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Articles 1 - 13 of 13

Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Examination Of Pseudomonas Fluorescence As A Recombinant Expression Host: Cloning, Expression, And Chromatography, Ahmed K.Ali Elmasheiti Dec 2016

Examination Of Pseudomonas Fluorescence As A Recombinant Expression Host: Cloning, Expression, And Chromatography, Ahmed K.Ali Elmasheiti

Theses and Dissertations

In an effort to expand the pool of bacterium useful for biotechnology applications, Pseudomonas fluorescens, a common gram negative microbe, was examined for its ability to function in a recombinant setting. P. fluorescens is ubiquitous in nature and was initially identified as a soil bacterium found in dirt and is typically associated with plant material. Past literature indicates that it shared characteristics common to Escherichia coli and Bacillus subtilis, including simple growth conditions and potential cloning vectors, providing motivation to look into both the upstream and downstream characteristics of this bacterium. First, it was demonstrated that P. fluorescens could be ...


Coarse-Grained Simulations Of The Self-Assembly Of Dna-Linked Gold Nanoparticle Building Blocks, Charles Wrightsman Armistead Dec 2016

Coarse-Grained Simulations Of The Self-Assembly Of Dna-Linked Gold Nanoparticle Building Blocks, Charles Wrightsman Armistead

Theses and Dissertations

The self-assembly of nanoparticles (NPs) of varying shape, size, and composition for the purpose of constructing useful nanoassemblies with tailored properties remains challenging. Although progress has been made to design anisotropic building blocks that exhibit the required control for the precise placement of various NPs within a defined arrangement, there still exists obstacles in the technology to maximize the programmability in the self-assembly of NP building blocks. Currently, the self-assembly of nanostructures involves much experimental trial and error. Computational modeling is a possible approach that could be utilized to facilitate the purposeful design of the self-assembly of NP building blocks ...


Membrane Chromatography For Bioseparations: Ligand Design And Optimization, Zizhao Liu Dec 2016

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

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 ...


Genotoxicity Of Graphene In Escherichia Coli, Ananya Sharma May 2016

Genotoxicity Of Graphene In Escherichia Coli, Ananya Sharma

Theses and Dissertations

Rapid advances in nanotechnology necessitate assessment of the safety of nanomaterials in the resulting products and applications. One key nanomaterial attracting much interest in many areas of science and technology is graphene. Graphene is a one atom thick carbon allotrope arranged in a two-dimensional honeycomb lattice. In addition to being extremely thin, graphene has several extraordinary physical properties such as its exceptional mechanical strength, thermal stability, and high electrical conductivity. Graphene itself is relatively chemically inert and therefore pristine graphene must undergo a process called functionalization, which is combination of chemical and physical treatments that change the properties of graphene ...


Investigation Of The Inherent Chemical, Structural, And Mechanical Attributes Of Bio-Engineered Composites Found In Nature: Alligator Gar’S Exoskeleton Fish Scales, Wayne Derald Hodo Dec 2015

Investigation Of The Inherent Chemical, Structural, And Mechanical Attributes Of Bio-Engineered Composites Found In Nature: Alligator Gar’S Exoskeleton Fish Scales, Wayne Derald Hodo

Theses and Dissertations

The U.S. Army has determined a huge cost savings of up to 51% can be accomplished by reducing the gross vehicle weight, for their personnel carrier, by 33%. To cut cost, composite materials are needed. Man-made composites can have superior material properties (high-strength, high-fracture toughness, and lightweight), but they are prone to delamination at the glued-layered interface. In contrast, fish scale is a natural composite that has the same material properties and, additionally, tend not to delaminate.

The focus of this study was to learn how nature integrates hard and soft materials at each length scale to form a ...


In Vivo Periodical Monitoring Of Immune Cell Infiltration In Response To Feathers And Intramuscular Injection Of Ionps Using The Pulp (Dermis) Of Growing Feathers As Test Site Tissue In Chickens, Olfat Taleb Alaamri Dec 2015

In Vivo Periodical Monitoring Of Immune Cell Infiltration In Response To Feathers And Intramuscular Injection Of Ionps Using The Pulp (Dermis) Of Growing Feathers As Test Site Tissue In Chickens, Olfat Taleb Alaamri

Theses and Dissertations

The applications of nanoparticles are growing, but little is known about their interactions with the immune system as most studies did not use suitable in vivo test systems. The purpose of these studies was to investigate the utility of chicken growing feathers as an in vivo test site for iron oxide nanoparticle (IONP) interactions with the immune system. The first objective of this study was to monitor leukocyte infiltration into the growing feather pulp upon the administration of IONP and IO-mIgG preparations. The second objective was to test the utility of IONPs as vaccine adjuvants by monitoring primary and memory ...


Characterization Of The Effects Of Bfgf And Gsk3Β Inhibitors On Embryoid Bodies Derived From Human Pluripotent Stem Cells, Jonathan Earls Jul 2015

Characterization Of The Effects Of Bfgf And Gsk3Β Inhibitors On Embryoid Bodies Derived From Human Pluripotent Stem Cells, Jonathan Earls

Theses and Dissertations

Embryoid body (EB) formation is a common first step in many human pluripotent stem cell (hPSC) differentiation protocols. Previous work suggests that EBs are sensitive to growth factor withdrawal if they are derived from hPSCs maintained in feeder independent media such as mTeSR1. To promote cell survival, EBs generated from mTeSR1-adapted hPSCs are sometimes cultured in a medium that contains basic fibroblast growth factor (bFGF), a trophic factor often used in hPSC cultures to maintain self-renewal. This distinguishes feeder independent hPSCs from feeder dependent hPSCs. The purpose of this study was to characterize the effects of bFGF as well as ...


Using Peptoids To Build Robust, Efficient Microarray Systems, Dhaval Sunil Shah Jul 2015

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

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.


Monodentate, Bidentate And Photocrosslinkable Thiol Ligands For Improving Aqueous Biocompatible Quantum Dots, Hiroko Takeuchi Dec 2013

Monodentate, Bidentate And Photocrosslinkable Thiol Ligands For Improving Aqueous Biocompatible Quantum Dots, Hiroko Takeuchi

Theses and Dissertations

Water-soluble Quantum Dots (QDs) are highly sensitive fluorescent probes that are often used to study biological species. One of the most common ways to render QDs water-soluble for such applications is to apply hydrophilic thiolated ligands to the QD surface. However, these ligands are labile and can be easily exchanged on the QD surface, which can severely limit their application. As one way to overcome this limitation while maintaining a small colloidal size of QDs, we developed a method to stabilize hydrophilic thiolated ligands on the surface of QDs through the formation of a crosslinked shell using a photocrosslinking approach ...


Expression, Production, And Purification Of Novel Therapeutic Proteins, Mckinzie Shea Fruchtl May 2013

Expression, Production, And Purification Of Novel Therapeutic Proteins, Mckinzie Shea Fruchtl

Theses and Dissertations

Interest in the production of recombinant proteins consisting of collagen binding domain (CBD) fused to a bioactive material has increased due to the targeting/attachment capabilities of CBD. For example, CBD fusions can be applied to the reversing of bone density loss and the repair of the eardrum, specifically, by choosing an appropriate fusion partner (parathyroid hormone or epidermal growth factor). The production of CBD fusions was examined using batch and fed-batch culturing of Escherichia coli to express the fusion proteins, and affinity chromatography to isolate the final product.

Different medium formulations, feeding strategies, and induction methods were tested in ...


The Geometry And Sensitivity Of Ion-Beam Sculpted Nanopores For Single Molecule Dna Analysis, Ryan Connor Rollings May 2013

The Geometry And Sensitivity Of Ion-Beam Sculpted Nanopores For Single Molecule Dna Analysis, Ryan Connor Rollings

Theses and Dissertations

In this dissertation, the relationship between the geometry of ion-beam sculpted solid-state nanopores and their ability to analyze single DNA molecules using resistive pulse sensing is investigated. To accomplish this, the three dimensional shape of the nanopore is determined using energy filtered and tomographic transmission electron microscopy. It is shown that this information enables the prediction of the ionic current passing through a voltage biased nanopore and improves the prediction of the magnitude of current drop signals when the nanopore interacts with single DNA molecules. The dimensional stability of nanopores in solution is monitored using this information and is improved ...


Capillary And Microchip Electrophoresis For The Monitoring Of Disease Causing Amyloid Proteins, Elizabeth Nancy Pryor Dec 2012

Capillary And Microchip Electrophoresis For The Monitoring Of Disease Causing Amyloid Proteins, Elizabeth Nancy Pryor

Theses and Dissertations

The detection of oligomers and aggregates formed by two amyloid proteins, insulin and amyloid-beta (AB), is of particular importance due to the role which these species play in Diabetes and Alzheimer's disease, respectively. However, existing techniques are limited in the ability to detect insulin and AB; oligomers due to the fact that these early aggregates are transient, present at low concentrations, and difficult to isolate. Improvements must be made to existing techniques or alternative techniques must be explored in order to identify and quantify the size of these oligomeric and aggregate species without disrupting their structure.

Capillary and microchip ...


Proteome Based Development Of Novel Affinity Tail For Immobilized Metal Affinity Chromatography And Hydrophobic Interaction Chromatography, Neha Tiwari Dec 2011

Proteome Based Development Of Novel Affinity Tail For Immobilized Metal Affinity Chromatography And Hydrophobic Interaction Chromatography, Neha Tiwari

Theses and Dissertations

At industrial scale, reducing the step in purification and recovery is desired; this not only decreases the cost but also increases the yield. Hydrophobic Interaction Chromatography (HIC) and Immobilized Metal Affinity Chromatography (IMAC) both are not harsh on biological structure or activity of proteins; also both the techniques are economical and therefore a suitable choice at industrial level. This dissertation comprises of three parts. Purpose for the Part I was to identify and characterize Escherichia coli proteins which display affinity towards both IMAC and Hydrophobic Interaction Chromatography (HIC). Co (II) IMAC was chosen as the primary capture step, followed by ...