Part I: Surface Modification of Magnetic Nanoparticles for the Recovery of Plasmid DNA and Exclusion of Endotoxins in Recombinant DNA Assays; Part II: Biosafety and Environmental Health Program Implementation Related to Manufacturing and Research of Medical Diagnostic Systems Focusing on Biopsy-Based Cancer Tests and Reagents

Autor: Young, Melissa A. L.
Rok vydání: 2016
Druh dokumentu: Text
Popis: Part I: Surface Modification of Magnetic Nanoparticles for the Recovery of Plasmid DNA and Exclusion of Endotoxins in Recombinant DNA Assays / Abstract: Plasmids are small circular double- stranded DNA molecules found naturally in bacterial cells. The genes that bacterial plasmids carry give the bacteria certain genetic advantages, such as being antibiotic resistant. As a bacterium divides the plasmids that it contains are copied and passed on to each daughter cell. Due to this unique nature of bacterial plasmids, scientists and researchers have employed the use of plasmids to clone, transfer and alter genes. Although quite useful for various molecular techniques, plasmid DNA which is destined for use in in vivo research pharmaceutical applications requires additional considerations due to other bacterial compounds bacterial cells possess. Endotoxin is a unique bacterial compound present in the outer membrane of Gramnegative bacteria. Endotoxin serves as a protective outer membrane permeability barrier, which gives the bacteria a defense mechanism in its environment. The endotoxin is released during the lysis of bacterial cells, as endotoxins can be hazardous and even deadly in certain concentrations. It is important that specialized techniques are utilized to purify plasmids. One particular technique developed by Ferradigm, a small Tucson, AZ based startup company, for purifying plasmids from bacterial cells, known as the Plasmid FlexPrep Kit, utilizes magnetic nanoparticles as a mode of extraction for plasmids. The Plasmid FlexPrep Kit used by Ferradigm involves basic DNA isolation chemistry. PEG (polyethylene glycol) is used to precipitate the plasmids out of solution, followed by a solid phase binding system of paramagnetic nanoparticles. This binding system allows the user to separate the bonded material out of solution using a magnet. Since plasmids (DNA) are negatively charged along with endotoxins there are issues of specificity. Thus for my internship project, I explored surface modification of the magnetic nanoparticles to increase specificity, which would in turn increase purity. Four different avenues were addressed including: affinity-ligand binding utilizing adsorption chemistry, nanoparticle metal typing, manufacture of magnetic nanoparticles utilizing a microwave in the presence of a proposed ligand, and surface coating of magnetic nanoparticles with humic acid to increase the ability of the nanoparticles to remain colloidal in solution. Results from these four different areas of investigation set the stage for further investigation aimed at the surface of magnetic nanoparticles and improving the specificity of the plasmid preparation assays. The concept of magnetic nanoparticle surface modification could enable scientist and researchers to specialize plasmid assays, and prepare plasmids for specific downstream in vivo experiments or potential pharmaceutical research applications, such as gene therapy.
Part II: Biosafety and Environmental Health Program Implementation Related to Manufacturing and Research of Medical Diagnostic Systems Focusing on Biopsy-Based Cancer Tests and Reagents / Abstract: Occupational and environmental health and safety are essential factors in the public health field. One’s occupation is the central activity for adults, a means to earn an income, and a center for the advancement of society. Safety in the workplace not only provides a secure environment for the employee, but it also reduces costs for the employer over time by diminishing work related injuries, illnesses and lost work time and production delays. Occupational health in turn plays a unique and important part in the environmental health as well. With improved health and safety in a workplace, workers are more conscience of how the hazardous material they dispose of affects themselves as well as their environmental surroundings. It is our duty as inhabitants of this planet, to protect our environmental surrounds. By understanding our ecological footprint contributed by daily work activities we can begin to improve our environmental health. In order to accomplish these goals health and safety programs specific to the occupation and local environmental concerns are necessary. Ventana Medical Systems Inc, is a cancer diagnostics company that specializes in the development and manufacture of primary stain, special stain, Immunohistochemistry (IHC), and in situ hybridization (ISH) staining platforms and reagents. These reagents and platforms are used in hospitals, clinical labs, and research facilities all over the world to diagnose and determine the proper treatment for specific cancers. With such high stakes resting on the quality and production of reagents and staining platforms, employee health and safety is crucial to maintain production of reliable products to customers. In response the Safety, Health and Environmental (SHE) department at Ventana is currently employing several different occupational and environmental health programs. Thus for my internship project I participated in compiling waste profiles, performing laboratory audits/walk-throughs, assisting with waste water and storm water profiling, and participating in risk assessments by attending risk assessment meetings and providing feedback. By setting these improved programs into place, SHE at Ventana can ensure that employees become acclimated to safety and health guidelines. Ventana Medical Systems Inc., has already shown progress in reducing waste by recycling, and increasing productivity by reducing employee injury in incident prone work areas. These programs will allow employees to conduct their research and manufacturing in safer and healthier work environments, as well as ensure that the company maintains its goals of improving the impact of business on the environment.
Databáze: Networked Digital Library of Theses & Dissertations
Externí odkaz: