Author:

Keywords:

aptamer; capillary electrophoresis; food allergen; lysozyme; systematic evolution of ligands by exponential enrichment, Science & Technology, Life Sciences & Biomedicine, Physical Sciences, Biochemistry & Molecular Biology, Chemistry, Multidisciplinary, Chemistry, aptamer, capillary electrophoresis, food allergen, lysozyme, systematic evolution of ligands by exponential enrichment, IN-VITRO SELECTION, LABEL-FREE, CAPILLARY-ELECTROPHORESIS, LIGAND-BINDING, HENS EGG, AFFINITY, PROTEIN, ACID, SPECTROSCOPY, MOLECULES, Aptamers, Nucleotide, Base Sequence, Binding Sites, DNA Primers, Egg White, Electrophoresis, Capillary, Enzyme-Linked Immunosorbent Assay, Fluorescence Polarization, Muramidase, Nucleic Acid Conformation, Polymerase Chain Reaction, Surface Plasmon Resonance, 0304 Medicinal and Biomolecular Chemistry, 0305 Organic Chemistry, 0307 Theoretical and Computational Chemistry, Organic Chemistry, 3404 Medicinal and biomolecular chemistry, 3405 Organic chemistry

Abstract:

We have selected aptamers binding to lysozyme from a DNA library using capillary electrophoresis-systematic evolution of ligands by exponential enrichment. During the selection process the dissociation constant of the ssDNA pool decreased from the micromolar to the low nanomolar range within five rounds of selection. The final aptamer had a dissociation constant of 2.8 ± 0.3 nM, 6.1 ± 0.5 nM, and 52.9 ± 9.1 nM as determined by fluorescence anisotropy, surface plasmon resonance and affinity capillary electrophoresis respectively. The aptamers were successfully challenged for specificity against other egg white proteins. The high affinity aptamers open up possibilities for the development of aptamer based food and medical diagnostics.