Engineering Commons^™

Bio-Separation Process Improvement Via Genomic Manipulation: Development Of Novel Strains For Use In Immobilized Metal Affinity Chromatography (Imac), Ryan Curtis Haley

Graduate Theses and Dissertations

The dissertation is comprised of three parts. Part I describes proteomic analysis of native bacterial proteins from Escherichia coli (E.coli) that bind during Immobilized Metal Affinity Chromatography (IMAC). Part II describes the value in exploiting proteome based data as a tool toward the design an E. coli expression strain that is particularly useful when Immobilized Metal Affinity Chromatography is employed as the initial capture step of a homologous protein purification process. Part III describes a methodology of chromosomal mapping of all contaminant gene products.

The objective of Part I was to identify all E. coli proteins that bind to Co(II), …

Valence-Length Correlations For Chemical Bonds From Atomic Orbital Exponents, F. D. Hardcastle

Journal of the Arkansas Academy of Science

Pauling’s empirical bond valence-length correlation has proven valuable because it offers a quick and convenient way of checking and evaluating molecular structures and determining oxidation states from measured bond lengths. In this study, a simplified quantum-mechanical approach was used to derive Pauling’s empirical bond valence-length relationship by considering overlap of hydrogen-like orbitals. An expression for the b “empirical” fitting parameter was derived in terms of atomic-orbital exponents. A new set of orbital exponents is presented using published atomic/covalent radii and a continuous function for the effective principal quantum. The b parameters calculated from the orbital exponents are consistent with bond …

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

Graduate 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 electrophoresis …

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

Graduate 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 …

Design And Fabrication Of Nanofluidic Systems For Biomolecule Characterizations, Orain Ansel Hibbert

Graduate Theses and Dissertations

Nanofluidic channel systems were designed and fabricated by combining MEMS microfabrication with AFM nanolithography. In the fabrication process flow, photolithography was first utilized to pattern microfluidic channels and reservoirs on a 4" Pyrex substrate. Subsequently, atomic force microscopy (AFM) based nanolithography was used to mechanically fabricate nanochannels to connect the microreservoirs which formed the inlet and outlet of the nanofluidic system. A Tap190 Diamond-Like Carbon (DLC) AFM tip with a force constant of 48 N/m and a radius of less than 15 nm was used as the nanolithography tool. The resultant nanochannel ranges from 20 to 80 µm in length …