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

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.

Grantor dc:publisher

Wake Forest University

Year dc:date.issued

2017

Author dc:creator

• Liu, Qian

• Binding constant

Language dc:language.iso

en

Handle dc:identifier.uri

http://hdl.handle.net/10339/82200

OAI identifier oai:identifier

oai:wakespace.lib.wfu.edu:10339/82200