Experimental data supporting DNA aptamer recognition of bilirubin and biliverdin for biosensing applications

This dataset supports the discovery and characterization of two DNA aptamers, Bvd4 and Brb7, that selectively bind to the bile pigments biliverdin and bilirubin, respectively—metabolites derived from heme degradation. These pigments are clinically relevant biomarkers associated with conditions such as neonatal jaundice, liver dysfunction, and hemolysis. The aptamers identified in this study may also serve as molecular switches for regulating gene expression. To support rigorous validation and reuse, the dataset includes raw experimental data obtained from: - Fluorescence titration assays using Thioflavin T (ThT) for binding curve generation. - Isothermal titration calorimetry (ITC) for thermodynamic profiling and dissociation constant (Kd) measurement. - Nuclear magnetic resonance (NMR) spectroscopy for structural validation and analysis of aptamer–ligand interactions. - UV–Vis spectroscopy for assessing pigment stability and monitoring bilirubin oxidation, - Sensor kinetic assays for evaluating real-time detection response and selectivity, and limit-of-detection (LOD) determination, The tightest biliverdin aptamer exhibited a Kd of 6 nM and an LOD of 0.7 nM. A separate aptamer showed specific binding to bilirubin with a Kd of 203 nM and an LOD of 47 nM. These values were confirmed using multiple orthogonal techniques. All experiments were conducted under controlled laboratory conditions at 25°C. Data are organized in tidy CSV format and correspond to Figures 3–8 and S1–S3 of the main article. Each file contains numeric measurements such as emission intensities, integrated heats, chemical shift data, and spectral information. This dataset facilitates further research into aptamer–target interactions, biosensor development, and nucleic acid-based sensing strategies for heme metabolites.