Persistent Link:
http://hdl.handle.net/10150/243914
Title:
Porous Phospholipid Nanoshells as Enzymes Delivery Agents
Author:
Comi, Troy J.
Issue Date:
May-2012
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
Diabetes is an epidemic in developed nations. Glucokinase (GK) is vital for glocose sensing, and is directly implicated in particular forms of diabetes. Studying pancreatic cells with altered GK activity would facilitate studies, but current methods for altering proteomes are lacking. Porous phospholipid nanoshells (PPNs) have traditionally been used as platforms for biologically derived nanosensors, though their biocompatibility and protease resistance well suits them as enzyme delivery agents. GK kinetics were investigated with an enzyme coupled reaction to determine the effect of encapsulation. It was determined that encapsulation increased the Hill coefficient by 5.8% and the S(0.5) by 1.8%. This small deviation may not be significant in physiological conditions. To observe a recovered function in cell lines upon reintroducing GK, constitutively expressed GK must first be knocked down with siRNA. As initial work toward an siRNA knockdown, immunoblotting conditions were optimized resulting in a detection limit below 10 ng of GK. Immunoblotting verified suspected constitutive expression of GK in INS-1 cell lines. While further investigation is necessary to demonstrate the utility of GK-containing PPNs for cell delivery, this thesis outlines the generation and characterization of this novel enzyme delivery platform.
Type:
text; Electronic Thesis
Degree Name:
B.S.
Degree Level:
bachelors
Degree Program:
Honors College; Biochemistry and Molecular Biophysics
Degree Grantor:
University of Arizona

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titlePorous Phospholipid Nanoshells as Enzymes Delivery Agentsen_US
dc.creatorComi, Troy J.en_US
dc.contributor.authorComi, Troy J.en_US
dc.date.issued2012-05-
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractDiabetes is an epidemic in developed nations. Glucokinase (GK) is vital for glocose sensing, and is directly implicated in particular forms of diabetes. Studying pancreatic cells with altered GK activity would facilitate studies, but current methods for altering proteomes are lacking. Porous phospholipid nanoshells (PPNs) have traditionally been used as platforms for biologically derived nanosensors, though their biocompatibility and protease resistance well suits them as enzyme delivery agents. GK kinetics were investigated with an enzyme coupled reaction to determine the effect of encapsulation. It was determined that encapsulation increased the Hill coefficient by 5.8% and the S(0.5) by 1.8%. This small deviation may not be significant in physiological conditions. To observe a recovered function in cell lines upon reintroducing GK, constitutively expressed GK must first be knocked down with siRNA. As initial work toward an siRNA knockdown, immunoblotting conditions were optimized resulting in a detection limit below 10 ng of GK. Immunoblotting verified suspected constitutive expression of GK in INS-1 cell lines. While further investigation is necessary to demonstrate the utility of GK-containing PPNs for cell delivery, this thesis outlines the generation and characterization of this novel enzyme delivery platform.en_US
dc.typetexten_US
dc.typeElectronic Thesisen_US
thesis.degree.nameB.S.en_US
thesis.degree.levelbachelorsen_US
thesis.degree.disciplineHonors Collegeen_US
thesis.degree.disciplineBiochemistry and Molecular Biophysicsen_US
thesis.degree.grantorUniversity of Arizonaen_US
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