Quantification via Inductively-Coupled Plasma Optical Emission Spectroscopy (ICP-OES) of the Cellular Internalization and Nuclear Localization of Gold Nanoparticles Passivated with BSA-SV40 Large T NLS Conjugates after Incubation with Human Cervical Cancer (HeLa) Cells

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dc.contributor.advisor Stefan Franzen, Committee Chair en_US
dc.contributor.advisor Edmond Bowden, Committee Member en_US
dc.contributor.advisor Kenneth Hanck, Committee Member en_US
dc.contributor.advisor Wayne Robarge, Committee Member en_US
dc.contributor.author Ryan, Joseph Anthony en_US
dc.date.accessioned 2010-04-02T18:53:58Z
dc.date.available 2010-04-02T18:53:58Z
dc.date.issued 2009-06-22 en_US
dc.identifier.other etd-06132009-145408 en_US
dc.identifier.uri http://www.lib.ncsu.edu/resolver/1840.16/4438
dc.description.abstract Rhodamine-labeled, cysteine-modified SV40 large T NLS peptide sequences were conjugated in varying amounts (~ 3 to 15 molar ratio) with bovine serum albuin (BSA) via the heterobifunctional linker succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC). These conjugates were then used to passivate nanomolar aliquots of citrate-coated gold nanoparticles of varying diameter (5, 10, 15, and 20 nm), and the stability of these nanoparticle complexes were evaluated with respect to: 1) amount of large T-BSA per nanoparticle (found to be stable under the following BSA:nanoparticle ratios: 5 nm diameter = 125:1, 10 nm diameter = 250:1, 15 nm diameter = 250:1, and 20 nm diameter = 500:1), 2) ionic strength of solution (critical coagulation concentration values above 0.85 M), and 3) temperature (found to be stable at 4, 25, and 37 degrees Celsius). A robust method was developed using inductively-coupled plasma optical emission spectroscopy (ICP-OES) as a means to quantify the internalization of nanoparticle complexes after incubation with human cervical cancer cells (HeLa) under many differing conditions. It was determined cellular internalization increased as a function of: 1) increasing amount of large T per nanoparticle complex, 2) longer incubation times, 3) temperature (which is to say incubations at 4 degrees Celsius afforded nearly no internalization), and 4) increasing nanoparticle diameter. Additionally, data from a pulse-chase experiment demonstrated that these nanoparticle complexes tend to remain associated with HeLa cells in similar concentration up to twelve hours after initial exposure. Lastly, a sub-cellular fractionation kit was used to extract nuclei from HeLa cells post-incubation with 5 nm diameter gold nanoparticle complexes. It was observed that nuclear localization of these nanoparticle complexes increased as a function of large T:nanoparticle ratio, but that resulting cell viability decreased dramatically at the highest large T:nanoparticle ratio. en_US
dc.rights I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dis sertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to NC State University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. en_US
dc.subject gold en_US
dc.subject nanoparticles en_US
dc.subject ICP-OES en_US
dc.subject HeLa en_US
dc.subject nuclear localization en_US
dc.subject delivery vector en_US
dc.title Quantification via Inductively-Coupled Plasma Optical Emission Spectroscopy (ICP-OES) of the Cellular Internalization and Nuclear Localization of Gold Nanoparticles Passivated with BSA-SV40 Large T NLS Conjugates after Incubation with Human Cervical Cancer (HeLa) Cells en_US
dc.degree.name PhD en_US
dc.degree.level dissertation en_US
dc.degree.discipline Chemistry en_US


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