Fig. 4. Characterization of RB aptamers.
A Aptamer candidates were converted to aptamer particles and incubated with 20 µM fluorescently labeled RA (orange) or RB (blue) prior to analysis via flow cytometry. Beads containing only the forward primer (FP) were used as a negative control. The y-axis shows median relative fluorescence unit (RFU) values and the error bars represent a single standard deviation of the three replicates. The raw data are overlayed on the bar plot as black dots. Source data are provided as a Source Data file. B Aptamer i-6 was expressed on beads and incubated with RA (orange) or RB (blue) prior to analysis via flow cytometry. The blue line represents a fitted Langmuir isotherm (KD = 29.5 µM ± 2.7 µM). Data are presented as median values and error bars represent the standard deviation of three replicate experiments. Source data are provided as a Source Data file. C Competitive binding assays for i-6 aptamer particles and 20 µM fluorescently-labeled RB alone or with 50 µM Man5 N-glycan (M5), 100 µM Lewis A (LeA) glycan, 10 mM mannose (Man), 10 mM GlcNAc, 50 µM M3, or 50 µM G0. The y-axis shows relative fluorescence normalized to the RB-only control. All experiments were conducted in triplicate and the median RFU values were recorded. The error bars represent a single standard deviation and the raw data points are overlayed on the bar plot as black dots. Source data are provided as a Source Data file. D Competitive binding assays conducted with i-6 aptamer particles and 20 µM of fluorescently labeled RB alone or with 20 µM of N-glycans containing five to nine mannose units (M5-M9). The y-axis shows relative fluorescence normalized to the RB-only control. All experiments were conducted in triplicate and the median RFU values were recorded. The error bars represent a single standard deviation and the raw data points are overlayed on the bar plot as black dots. Source data are provided as a Source Data file. E Structures of the molecules used in the competition assays.