title: Monitoring aptamer-protein interactions using tunable resistive pulse sensing.
abstract: Aptamers are short single-stranded pieces of DNA or RNA capable of binding to analytes with specificity and high affinity. Due to their comparable selectivity, stability, and cost, over the last two decades, aptamers have started to challenge antibodies in their use on many technology platforms. The binding event often leads to changes in the aptamer's secondary and tertiary structure; monitoring such changes has led to the creation of many new analytical sensors. Here, we demonstrate the use of a tunable resistive pulse sensing (TRPS) technology to monitor the interaction between several DNA aptamers and their target, thrombin. We immobilized the aptamers onto the surface of superparamagnetic beads, prior to their incubation with the thrombin protein. The protein binding to the aptamer caused a conformational change resulting in the shielding of the polyanion backbone; this was monitored by a change in the translocation time and pulse frequency of the particles traversing the pore. This signal was sensitive enough to allow the tagless detection of thrombin down to nanomolar levels. We further demonstrate the power of TRPS by performing real time detection and characterization of the aptamer-target interaction and measuring the association rates of the thrombin protein to the aptamer sequences.
authors.affiliation: Department of Chemistry, Centre for Analytical Science, Loughborough University , Loughborough, Leicestershire LE11 3TU, United Kingdom.
authors.forename: Emily R
authors.initials: ER
authors.lastname: Billinge
authors.forename: Murray
authors.initials: M
authors.lastname: Broom
authors.forename: Mark
authors.initials: M
authors.lastname: Platt
mesh_terms: D052157:Aptamers, Nucleotide / Q000138:chemical synthesis / Q000737:chemistry*; D015374:Biosensing Techniques; D055664:Electrochemical Techniques; D006801:Humans; D007700:Kinetics; D059346:Magnets; D053758:Nanoparticles / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D012680:Sensitivity and Specificity; D012996:Solutions; D013917:Thrombin / Q000032:analysis*
chemical_list: D052157:Aptamers, Nucleotide; D012996:Solutions; D013917:Thrombin