Faculty Mentor Information
Dr. Daniel Fologea (Mentor), Boise State University
Abstract
This work is focused on investigating the utilization of aptamers in conjunction with the Kinetics Exclusion Assay (KinExA) technology for detection and quantification of the antibiotic ampicillin in solutions. To achieve the scientific objectives of this project, I exploited the strong affinity of biotin for streptavidin and functionalized microscopic beads to be utilized as capture elements for fluorescent, free ampicillin aptamers left in equilibrated solutions. To capture single stranded DNA aptamers, the last layer immobilized on the beads comprised single stranded DNA molecules complementary to the aptamer’s sequence. Equilibrated mixtures of a fixed aptamer concentration and variable ampicillin concentrations have been flown over the beads and assessed with the KinExA 4000 instrument to evaluate the fraction of free aptamers in solutions. The experimental data have been plotted and analyzed to determine the affinity of the aptamers for ampicillin, estimate the range of quantifiable ampicillin concentrations, and measure the ampicillin concentrations in three mock samples. The data demonstrate that ampicillin can be accurately detected and quantified by employing the KinExA technology; the proposed methodology may be adapted for quantification of other molecules of interest, including disease biomarkers and environmental pollutants.
Quantification of Ampicillin Through Kinetic Exclusion Assay (KinExA) Technology
This work is focused on investigating the utilization of aptamers in conjunction with the Kinetics Exclusion Assay (KinExA) technology for detection and quantification of the antibiotic ampicillin in solutions. To achieve the scientific objectives of this project, I exploited the strong affinity of biotin for streptavidin and functionalized microscopic beads to be utilized as capture elements for fluorescent, free ampicillin aptamers left in equilibrated solutions. To capture single stranded DNA aptamers, the last layer immobilized on the beads comprised single stranded DNA molecules complementary to the aptamer’s sequence. Equilibrated mixtures of a fixed aptamer concentration and variable ampicillin concentrations have been flown over the beads and assessed with the KinExA 4000 instrument to evaluate the fraction of free aptamers in solutions. The experimental data have been plotted and analyzed to determine the affinity of the aptamers for ampicillin, estimate the range of quantifiable ampicillin concentrations, and measure the ampicillin concentrations in three mock samples. The data demonstrate that ampicillin can be accurately detected and quantified by employing the KinExA technology; the proposed methodology may be adapted for quantification of other molecules of interest, including disease biomarkers and environmental pollutants.
Comments
Author name variant: Zhiyu Li