Aptamer Selection, Characterization, 3D Structure Simulation And Target Identification
After mathematical analysis of the sequences obtained by Next Generation Sequencing, up to 10 aptamers are selected, which are then synthesized chemically. Their specificity and affinity for the target is assessed using flow cytometry and microscopy. The experiments on the binding of aptamers to positive and negative targets are performed in several replicates and are statistically processed. As a result of the analysis, 1-3 aptamers with the best characteristics are selected.
The dissociation constant (Kd) allows one to evaluate at what concentrations the target and aptamer enter the equilibrium state. This value allows one to indirectly assess the optimal concentration of aptamers for binding. Experiments to evaluate Kd of aptamers and a positive target are performed by flow cytometry.
Aptamers affinity for target tissues, cells, bacteria or fungi is estimated using flow cytometry.
Aptamers specificity for negative tissues, cells, bacteria or fungi is estimated using flow cytometry and laser scanning microscopy.
The dissociation constant (Kd) of an aptamer and it's target is estimated using flow cytometry.