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http://hdl.handle.net/11375/17354
Title: | Advances in the Use of Sol-Gel-Derived Microarrays as an Assay and Detection Platform |
Authors: | Lebert, Julie M. |
Advisor: | Brennan, John D. |
Department: | Chemical Biology |
Keywords: | chemical biology, sol-gel-derived, microassay, detection |
Publication Date: | Aug-2009 |
Abstract: | The use of sol-gel immobilization in the fabrication of microarrays is a relatively new approach that has shown potential to become a leading methodology in this field. However, there are a limited number of assay systems that have been reported using this method. Furthermore, methods to produce high-density sol-gel-derived microarrays have not been reported. Herein, two novel assays utilizing sol-gel-derived microarrays are presented. In the first case, the solid phase of sol-gel-derived microarrays was employed as a detection platform for monitoring the activity of glycogen synthase kinase 3-β (GSK3β) in solution using a phosphospecific stain. Using this assay format, the ability to detect hyperphosphorylated product over the pre-phosphorylated substrate was demonstrated and a z' value of 0.49 was obtained, indicating amenability to small molecule screening. Secondly, a fluorogenic assay for acetylcholinesterase (AChE) was development that is compatibility with sol-gel derived microarrays and standard imaging instrumentation. A thiol-reactive fluorogenic dye, typically used for detection of thiolated oligonucleotides, was successfully used to monitor AChE activity both in solution and in silica. Further, a functional sol-gel-derived AChE microarray was fabricated and activity on array was detected. We have also reported on the optimization of materials for the fabrication of high-density kinase microarrays using sol-gel immobilization. By employing a directed criteria-based screen, optimal materials were quickly and efficiently identified. Two materials, 1.5SS/1PV A/Glycerol and 0.25DGS, were identified as the optimal materials for fabrication of sol-gel-derived functional microarrays. |
URI: | http://hdl.handle.net/11375/17354 |
Appears in Collections: | Open Access Dissertations and Theses |
Files in This Item:
File | Description | Size | Format | |
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Lebert_Julie_M_2009Aug_MSc.pdf | 10.06 MB | Adobe PDF | View/Open |
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