2D layout of bead based thioaptamer/aptamer selection platform for therapeutics and diagnostics

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dc.contributor.advisor Bruce A . Luxon en_US
dc.contributor.committeeMember William D . Willis en_US
dc.contributor.committeeMember L .Lilian Chan en_US
dc.creator Xu Zhao en_US
dc.date.accessioned 2011 -12 -20T16 :05 :04Z
dc.date.accessioned 2014 -02 -19T22 :05 :26Z
dc.date.available 2008 -06 -17 en_US
dc.date.available 2011 -12 -20T16 :05 :04Z
dc.date.available 2014 -02 -19T22 :05 :26Z
dc.date.created 2007 -07 -25 en_US
dc.date.issued 2007 -07 -20 en_US
dc.identifier.other etd -07252007 -082849 en_US
dc.identifier.uri http : / /hdl .handle .net /2152 .3 /187
dc.description.abstract Nucleic acid research has expanded the way we can intervene with biological systems . Especially , oligonucleotide agents (ODN or aptamers ) are believed to affect cell function via complementary recognition or binding to specific proteins by forming tertiary structure . This opens new ways in therapeutics and diagnostics . The phosphoro - mono - /di - thioate substitutions in the backbone (termed “thioaptamer” ) grants ODN nuclease resistance and higher binding affinity . \r \nA bead -based combinatorial library , in which every bead contains a unique species of aptamers , provides a promising platform for selection of aptamers and thioaptamers . To successfully screen the bead -based library , 2D layout of beads in gel and on bead screening model is proposed . To develop the 2D layout of beads and its corresponding functional assays , a model system is first established : NF -kappa B proteins were expressed , purified and characterized . Thioaptamer XBY6 , which specifically targets NF -kappa B protein , and its natural origin , I -kappa B were synthesized and verified . Thioaptamer purification using FPLC and HPLC was also investigated , and several 5’ -funtionalized thioaptamers were successfully purified . Electrophoretic mobility shifting assay (EMSA ) has been used to verify XBY6 binding , and ELISA assay has been used to verify I -kappa B binding towards human recombinant NF -kappa B protein . Preliminary study of bead in 2D gel showed applicability of bead -based selection and thus on bead functional assays were developed . Both double strand one species library with I -kappa B sequence and a 212=4096 different species beads library were constructed and verified . The library was then tested using on -bead EMSA like assay and ELISA assay . Both assays showed encouraging results for 2D layout selection and further enhancement of visualization (signal /noise improvement ) is discussed . \r \nThe project suggested that 2D layout of beads in gel (PAGE ) is well suited for parallel high -throughput selection of thioaptamers and aptamers , thus paving a new way for drug discovery and future therapeutics and diagnostics . \r \n en_US
dc.format.medium electronic en_US
dc.language.iso eng en_US
dc.rights Copyright © is held by the author . Presentation of this material on the TDL web site by The University of Texas Medical Branch at Galveston was made possible under a limited license grant from the author who has retained all copyrights in the works . en_US
dc.subject Thioaptamer XBY6 en_US
dc.subject nucleic acid en_US
dc.subject Nuclear Factor Kappa B en_US
dc.subject drug development en_US
dc.subject combinatorial chemistry en_US
dc.subject bead -based selection en_US
dc.subject Aptamer en_US
dc.title 2D layout of bead based thioaptamer /aptamer selection platform for therapeutics and diagnostics en_US
dc.type.genre thesis en_US
dc.type.material text en_US
thesis.degree.name Master of Science en_US
thesis.degree.level Master en_US
thesis.degree.grantor The University of Texas Medical Branch en_US
thesis.degree.department Human Biological Chemistry and Genetics en_US

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2D layout of bead based thioaptamer/aptamer selection platform for therapeutics and diagnostics. The University of Texas Medical Branch. Available electronically from http : / /hdl .handle .net /2152 .3 /187 .

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