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dc.contributor.authorHolub, Douglas F.
dc.descriptionDissertation (Ph.D.)-- University of Kansas, Chemistry, 2005.en_US
dc.description.abstractAptamers have been touted as viable complements to antibodies as molecular recognition elements in diagnostic assays. Several studies have demonstrated aptamer applicability in both homogeneous and heterogeneous assay formats. However, very few studies have been performed to investigate the parameters involving effect immobilization of aptamers, the analyte-binding kinetics of attached aptamers, and the optimal assay format for aptamers. These studies utilized the well-characterized anti-thrombin DNA aptamer. Investigations were conducted in order to ascertain effective methods of immobilization of the aptamer to silica and gold, the maximum packing density for which 100% of the aptamer activity is retained, and the binding kinetics of the immobilized aptamer. The attachment studies found that a short linker extending from the aptamer is essential for covalent attachment, the aptamer tertiary structure and cation dependence can influence the effectiveness of a covalent attachment, and affinity attachment methodologies are reliable for aptamer immobilization. In addition, the optimal packing density for active aptamers was determined to be similar to the reported optimal packing density of singlestranded oligonucleotides used for hybridization assays. The kinetic studies for the binding of the immobilized aptamer to its cognate ligand showed that the binding kinetics of the aptamer are unaffected by either covalent or affinity attachment to a substrate, the association and dissociation binding rates for this aptamer are very fast, and the association rate is diffusion-limited. These results indicate that aptamers may be well-suited to use in miniaturized, real-time, continuous-monitoring biosensors.en_US
dc.publisherUniversity of Kansasen_US
dc.rightsThis item is protected by copyright and unless otherwise specified the copyright of this thesis/dissertation is held by the author.en_US
dc.subjectPure sciencesen_US
dc.subjectBiological sciencesen_US
dc.subjectDNA aptameren_US
dc.subjectPacking densityen_US
dc.titleImmobilization Of The Anti-Thrombin DNA Aptamer: Attachment, Optimal Packing Density And Kineticsen_US
kusw.oanotes08/24/2022: Added to KU ScholarWorks with the permission of the author: From: "Holub, Douglas F." <> Date: August 25, 2022 at 7:00:25 AM CDT To: "Wilson, George S." <> Subject: Re: ScholarWorks Hello Dr. Wilson,

Thank you for contacting me. I would definitely like to have my dissertation published in ScholarWorks. Please let me know if I need to do anything else to facilitate this.

Cleaning out your office must be difficult- what with all of the memories associated with the material. I know that I get a little melancholy at the end of each semester when leaving my office for the last time.

Wishing you and Dr. Beverly the best,


Douglas F. Holub, PhD Associate Professor of Chemistry 616 Evenden Tower SUNY Delhi Delhi, NY 13753 785-817-7649 Zoom Meeting Room:

Your email is important to me. I aim to respond within the day received, but please note that it may take 24-48 hours (two business days) to respond to emails. ________________________________________ From: Wilson, George S. <> Sent: Wednesday, August 24, 2022 5:13 PM To: Holub, Douglas F. <> Subject: ScholarWorks Hi Doug: I assume that this address works for you. I am in the process of cleaning out my office in Malott and find that I have a large number of dissertations there. You may also know that in the interest of Open Access KU has made possible a library of digitized dissertations which are now freely accessible to anyone. Consequently I am trying to get all the KU dissertations into ScholarWorks. For many of the other students such as Raeann where the rules are somewhat different this is now being accomplished. Your dissertation was submitted to University Microfilms. If you would like to have your dissertation concluded in ScholarWorks, then I need an e-mail giving KU the permission to so publish it. You have the present copyright and that is maintained. Students who got their degrees before 2005 were also offered the opportunity to get a complete hard copy of their dissertation. That is not possible for you. I don’t have a complete copy of your dissertation so I cannot sent it to you. The library has a copy which they will retain so if you want a complete copy you will need to download the electronic form. If you would like your dissertation included in ScholarWorks, send me an e-amil indicating your accord. I hope all goes well with you. Best, George

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