Biotechnology Commons^™

Mammalian Cell Line Development Platform For Recombinant Protein Production: Expanding The Protein Expression Toolbox For Research And Drug Discovery Applications, Elizabeth Del Greco

Dissertations, Masters Theses, Capstones, and Culminating Projects

Recombinant proteins have revolutionized the biomedical industry, providing therapeutics for life-threatening diseases and protein reagents for research applications. BioMarin Pharmaceutical Inc. develops recombinant protein therapeutics to treat rare diseases including lysosomal storage disorders (LSDs), a group of about 50 individually rare disorders together affecting 1 in 8,000 live births. With an increase in the number of novel therapeutics in our drug discovery pipeline, there is a high demand to produce a variety of recombinant proteins for early-stage drug development projects. In order to equip our protein production process with the tools and capability for diverse protein expression, it is valuable …

Understanding The Mechanisms Of Insecticide Resistance In Phlebotomus Papatasi And Lutzoymia Longipalpis Sand Flies (Diptera: Psychodidae: Phlebotominae), David Denlinger

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Sand flies, like mosquitoes, ticks, fleas, and lice, transmit pathogens that cause disease in humans. Leishmaniasis, caused by pathogens transmitted by sand flies, kills tens of thousands of people every year. Insecticides have been used to control sand flies, but there is evidence of insecticide resistance in populations of sand flies around the world. The goal of this dissertation was to develop tools to maintain sand flies in the laboratory, develop the ability to identify insecticide-resistant populations of sand flies, and to investigate the genetic mechanisms of how sand flies become resistant to insecticides. I began by comparing live animal …

An Assessment Of Seven New Grape Varieties And A Study Of Grapevine Vein Clearing Virus In Native Vitaceae Plants, Kaylie A. Austin

Graduate Theses/Dissertations

One of the most effective methods of reducing pesticide application in vineyards is breeding hybrid varieties for disease resistance. The new varieties must be assessed for disease resistance and viticultural traits. This thesis focuses on seven new varieties from a cross of ‘Norton’ and ‘Cabernet Sauvignon’. ‘Norton’ was chosen because of its disease resistance and cold hardiness. ‘Cabernet Sauvignon’ was selected because of its high-quality wine making berries. The cross was made in 2005, seven were selected in 2011, and evaluation began in 2016. Using Botrytis cinerea as a model pathogen, resistance was measured with an incidence rate and severity …

Dynamic Protein Movement Involved In Photoreceptors, Daisy Spoer

Renée Crown University Honors Thesis Projects - All

Ciliopathies are a class of genetic diseases which affect the function of primary cilium within multiple organs throughout the body. Although ciliopathies are known to present high morbidity and mortality rates, there are no current cures and only a limited amount of treatments. A universal function of cilia is to regulate cellular activities through the transduction of signals. Disruption of the localization of proteins following the reception of a stimuli can be responsible for the phenotypic effects of dysfunctioning cilia in ciliopathies. In order to address the pathogenesis, we must first understand how proteins move within the cilia, so that …

Estimating Autoantibody Signatures To Detect Autoimmune Disease Patient Subsets, Zhenke Wu, Livia Casciola-Rosen, Ami A. Shah, Antony Rosen, Scott L. Zeger

Johns Hopkins University, Dept. of Biostatistics Working Papers

Autoimmune diseases are characterized by highly specific immune responses against molecules in self-tissues. Different autoimmune diseases are characterized by distinct immune responses, making autoantibodies useful for diagnosis and prediction. In many diseases, the targets of autoantibodies are incompletely defined. Although the technologies for autoantibody discovery have advanced dramatically over the past decade, each of these techniques generates hundreds of possibilities, which are onerous and expensive to validate. We set out to establish a method to greatly simplify autoantibody discovery, using a pre-filtering step to define subgroups with similar specificities based on migration of labeled, immunoprecipitated proteins on sodium dodecyl sulfate …

Validation Of The Pre-B Cell Receptor As A Therapeutic Target In B Cell Precursor Acute Lymphoblastic Leukemia, Michael Frank Erasmus

Biomedical Sciences ETDs

This dissertation is built upon the fundamental idea that the pre-B cell receptor (pre-BCR) is important to leukemia cell survival and a logical therapeutic target in B cell precursor acute lymphoblastic leukemia (BCP-ALL). The pre-BCR is expressed early at a specific stage during B cell development where it plays a central role in survival of healthy B lymphocytes. This receptor is composed of the membrane heavy chain (mIgμ) associated with surrogate light chain components, 5 and VpreB. Through the use of advanced imaging modalities, in particular two-color single particle tracking (SPT), we showed that pre-BCRs formed transient, homotypic interactions. These …

Crispr/Cas9 Genome Editing System And Its Use In Infectious Disease, Dustin Daws

Thinking Matters Symposium Archive

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is an exciting new genetic engineering technology that was found in the chromosomes of certain bacteria and archaea. Bacteriophage are viruses that insert their DNA and hijack the host cell’s machinery to make new phage that can go on to infect new cells. Some microorganisms use CRISPR as a defense mechanism to disrupt the bacteriophage DNA after it is inserted into the cell. CRISPR/Cas9 uses genome editing as a means to alter very specific sections of a foreign genome. CRISPR works with a RNA-guided DNA endonuclease called Cas9. This enzyme can locate and …

A Study Of Methods For Detecting Gene Expression Of The Sterol Biosynthetic Pathway In The Microalgae Strain Heterosigma Akashiwo, Jessica C. Mayhew, Marcia Ackerman, Mike Lomas

Thinking Matters Symposium Archive

Biotechnological applications for various microalgae strains are beginning to bloom. There is interest in these microbes for their production of useful compounds such as phytosterols and lipids which can be produced in mass culture for large scale manufacture in the food, pharmaceutical, and biofuel industries (Romano et al., 2016). To this end, we are using the gold-brown microalgae, Heterosigma akashiwo (CCMP1680), a known producer of phytosterols, to test a molecular method for quick and easy measurements of production of the phytosterol compound sitosterol, which has potential as an anti- inflammatory agent. (Giner et al., 2008).

Stem Cell Derived Osteoprogenitors And Their Role In Bone Repair Using Morphogenic Activators, George Washington Brownridge Iii, Sylvanna Islas, Angelina Miller, Warren Hoeffler

Student Research Posters

Bone constantly cycles through a dynamic process of breakdown and remodeling. Osteoblasts are the specialized mesenchymal stem cells that have a major role in bone formation and the remodeling process whereas their counterpart osteoclasts, handle bone resorption. Embryonic stem cells can be partially differentiated into Progenitor cells, and we worked with #18, a candidate for being an osteoprogenitor that has the potential to respond to morphogenic activators. In the case of bone remodeling, TGF-β 2, BMP-2 and an abundance of CA++ have been shown to be potential activators of differentiation into osteoblasts. Eight different trials were conducted with the cells …

Disappearance And Appearance Of An Indigestible Marker In Feces From Growing Pigs As Affected By Previous- And Current-Diet Composition, Brandy M. Jacobs, John F. Patience, Merlin D. Lindemann, Kenneth J. Stalder, Brian J. Kerr

Animal and Food Sciences Faculty Publications

Background: Indigestible markers are commonly utilized in digestion studies, but the complete disappearance or maximum appearance of a marker in feces can be affected by diet composition, feed intake, or an animal’s BW. The objectives of this study were to determine the impact of previous (Phase 1, P1) and current- (Phase 2, P2) diet composition on marker disappearance (Cr) and appearance (Ti) in pigs fed 3 diets differing in NDF content.

Results: When pigs were maintained on the 25.1, 72.5, and 125.0 g/kg NDF diets, it took 5.1, 4.1, and 2.5 d, respectively, for Cr levels to decrease below the …

Oral History Conversation With Lydell Aaron (Beartha), Alexander Z. Bulkin, Kari Schattinger, Kate L. Gardner, Charbel Mouannes, Danielle Silvaggio

Philosophy: All Student Work

Oral History Interview conducted with Lydell Aaron. He founded Beartha Systems which builds scalable and smart systems that harness energy and resources for small communities.

Microbal Strans And Methods Of Making And Using, Nicole Roswitha Buan Murphy, Jennifer Catlett

Department of Biochemistry: Faculty Publications

Microbial strains are provided, as are methods of making and using Such microbial strains.

P02. Production Of A Protein Subunit Vaccine For Mannheimia Haemolytica In Lettuce Chloroplasts, Coby K. Martin

Western Research Forum

The cattle industry worldwide is ravaged by bovine respiratory disease (BRD), a bacterial disease caused by Mannheimia haemolytica. We have designed a chimeric protein subunit vaccine against M. haemolytica based on recent evidence demonstrating the protective potential of antigens against a virulence factor, leukotoxin, in addition to a surface lipoprotein. Plant-based production of this protein vaccine provides a safe and inexpensive alternative to traditional production methods. Plant-base production also supports the use of an edible vaccine that will deliver antigens to pharyngeal tissues to provide local immunization against M. haemolytica to prior to its progression into the lungs. Chloroplasts …

P08. Unravelling Organelle Genome Evolution Architecture Using Rna-Sequencing Data, Matheus Sanita Lima, David Roy Smith

Western Research Forum

Background: Mitochondria genomes vary from 11 Mb to 6 kb, while plastids can vary from 1 Mb to 30 kb. Non-coding DNA accounts for most of this size variation, but the mechanistic and evolutionary reasons for that are still unknown. Next generation sequencing has generated unprecedented amounts of genomic and transcriptomic data that can be used for organelle genome evolution studies. However, most of these data is used only for the study of cell nucleus. Therefore, I decided to use these untapped data source to investigate the transcription of organelle genomes in plastid-bearing protists.

Methods: I mapped the transcriptomes over …

Repository Review: Nasa Pubspace, Laura Pope Robbins

Publications

Laura Pope Robbins is an advisor for The Charleston Advisor, and the review below is included in Scholarly Commons with the permission of the publisher.

PubSpace is a repository of full-text peer-reviewed articles resulting from NASA-funded research going back to 1961. The National Center for Biotechnology Information has integrated PubSpace into PubMed Central, a freely available repository of medical research. Using an established, stable, government-hosted platform for PubSpace seems to make sense. However, the lack of clear branding, an uneven application of phrase searching, and a missing thesaurus of NASA terminology highlights that this integration may not be the best.

Pneb193-Derived Suicide Plasmids For Gene Deletion And Protein Expression In The Methane-Producing Archaeon, Methanosarcina Acetivorans, Mitchell T. Shea, Mary E. Walter, Nikolas Duszenko, Anne-Lise Ducluzeau, Jared Aldridge, Shannon K. King, Nicole R. Buan

Department of Biochemistry: Faculty Publications

Gene deletion and protein expression are cornerstone procedures for studying metabolism in any organism, including methane-producing archaea (methanogens). Methanogens produce coenzymes and cofactors not found in most bacteria, therefore it is sometimes necessary to express and purify methanogen proteins from the natural host. Protein expression in the native organism is also useful when studying post-translational modifications and their effect on gene expression or enzyme activity. We have created several new suicide plasmids to complement existing genetic tools for use in the methanogen, Methanosarcina acetivorans. The new plasmids are derived from the commercially available E. coli plasmid, pNEB193, and cannot replicate …

The Role Of Estrogen In Mouse Gastric Stem Cell Homeostasis, Aysha Mohamed Yusuf Alkaabi

Theses

To maintain cellular homeostasis, the epithelial lining of the stomach wall continuously fluctuates between cellular proliferation, differentiation, and apoptosis. A key player in this process is the gastric stem cell (GSC). GSCs are located in the isthmus region of the corpus gastric gland and have the potential to proliferate and differentiate. Although several pathways have been identified to regulate stem cell role in several body tissues, little is known about controlling GSC homeostasis. This project aims to study the role of estrogen (E2) in GSC homeostasis using the well-established mouse gastric epithelial progenitor (mGEP) cell line. Our data showed that …

Towards The Development Of A Sustainable Soya Bean-Based Feedstock For Aquaculture, Hyunwoo Park, Steven Weier, Fareha Razvi, Pamela A. Peña, Neil A. Sims, Jennica Lowell, Cory Hungate, Karma Kissinger, Gavin Key, Paul Fraser, Jonathan Napier, Edgar B. Cahoon, Thomas Clemente

Center for Plant Science Innovation: Faculty Publications

Soya bean (Glycine max (L.) Merr.) is sought after for both its oil and protein components. Genetic approaches to add value to either component are ongoing efforts in soya bean breeding and molecular biology programmes. The former is the primary vegetable oil consumed in the world. Hence, its primary usage is in direct human consumption. As a means to increase its utility in feed applications, thereby expanding the market of soya bean coproducts, we investigated the simultaneous displacement of marine ingredients in aquafeeds with soya bean-based protein and a high Omega-3 fatty acid soya bean oil, enriched with alpha-linolenic …

Engineering Fret Biosensors For Microrna Presence/Absence Analysis, Nicholas E. Larkey, Sean M. Burrows

Biomedical Engineering Western Regional Conference

No abstract provided.

Portable, Powerless Automation Of Valve Actuation For Microfluidic Large-Scale Integration Technology, Andrew Schmidt Mr., Matt Fitzgerald Mr., Emre Araci Mr., Connor Mcloughlin Mr.

Biomedical Engineering Western Regional Conference

Portable, powerless automation of valve actuation for microfluidic large-scale integration technology