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CAPILLARY ELECTROPHORESIS ANALYSIS OF SINGLE-STRANDED DNA APTAMERS AND THEIR INTERACTIONS WITH PROTEIN TARGETS AND CARBON DOTS

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abstract
Aptamers are short, single-stranded oligonucleotides, which fold into particular 3-dimensional structures permitting high-affinity binding to target molecules. In the first part of this work, capillary electrophoresis experiments were conducted to determine the dissociation constant of a unique FITC-labeled single-stranded DNA and anti-FITC antibody system in free solution environment using the method of non-equilibrium capillary electrophoresis of non-equilibrium mixtures (NECEEM). The free-solution binding constant was 316 ± 17 nM. The second part of this work was intended to shed light on the nature of aptamer interactions with nanomaterials used in an attempt to improve the resolution of aptamers bound to small molecular weight targets such as drugs and drug metabolites. Capillary electrophoresis with UV/Vis absorbance detection and laser-induced fluorescence detection (CE-UV and CE-LIF) was used to explore whether or not carbon dots (CDs) could significantly change the mobility of ssDNA samples upon noncovalent interaction. Preliminary observations revealed substantial interactions between carbon dots and ssDNA, as evidenced by changes in fluorescence emission of labeled DNA samples, but further studies are needed to prove whether or not CDs can significantly modify the electrophoretic mobility of ssDNA probes for analytical purposes.
subject
Binding constant
Capillary electrophoresis
Carbon dots
DNA aptamers
Fluorescence detection
NECEEM
contributor
Liu, Qian (author)
Colyer, Christa L. (committee chair)
Hinze, Willie (committee member)
Wright, Marcus (committee member)
date
2017-06-15T08:35:57Z (accessioned)
2019-06-14T08:30:14Z (available)
2017 (issued)
degree
Chemistry (discipline)
embargo
2019-06-14 (terms)
identifier
http://hdl.handle.net/10339/82200 (uri)
language
en (iso)
publisher
Wake Forest University
title
CAPILLARY ELECTROPHORESIS ANALYSIS OF SINGLE-STRANDED DNA APTAMERS AND THEIR INTERACTIONS WITH PROTEIN TARGETS AND CARBON DOTS
type
Thesis

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