Table 1.
Summary of the instrumental methods, buffer conditions and binding models previously used to determine binding constants of the thrombin aptamers.
| Aptamer | Binding Constant | Method | Buffer Conditions | Model |
|---|---|---|---|---|
| 15mer | EC50 = 25nM (Bock et al, 1992) |
Nitrocellulose Filter Binding | 20mM Tris-acetate, pH 7.4, 140 mM NaCl, 5mM KCl, 1mM CaCl2, 1mM MgCl2 | N/A |
| Kd = 54.911nM (Lin et al, 2011) |
Surface Plasmon Resonance | 10mM Tris-HCl, 5mM KCl, 1mM MgCl2, 1mM CaCl2, 50mM NaCl, pH 7.4 | 1:1 from kinetics | |
| Kd = 20nM (Huang et al, 2004) |
Affinity Probe Capillary Electrophoresis | 10mM Tris-HCl, 15mM KCl, pH 8.4 | NECEEM | |
| Kd = 450nM (German et al, 1998) |
Affinity Probe Capillary Electrophoresis | 5mM Na2HPO4, 5mM KH2PO4, 2mM MgCl2, pH 8.2 | Simple Isotherm | |
| Kd = 240±16nM (Berezovski et al, 2003) |
Affinity Probe Capillary Electrophoresis | 20mM Tris-HCL, pH 8.3, 5mM KCl, 1mM MgCl2 | NECEEM | |
| Kd = 43nM (Gong et al, 2008) |
MicroChip Affinity Capillary Electrophoresis | 25mM Tris, 192mM 5mM HCl | NECEEM | |
| Kb = 3±1e6 M−1 (Pagano et al, 2008) |
Isothermal Titration Calorimetry | 10 mM potassium phosphate, 70 mM KCl, 0.1 mM EDTA at pH 7.0 | Unspecified 1 aptamer:2 protein model | |
| Kd = 30±19nM (Baaske et al, 2010) |
Optical Thermophoresis | 20mM Tris-acetate, pH 7.4, 140 mM NaCl, 5mM KCl, 1mM CaCl2, 1mM MgCl2 | N/A | |
| EC50 = 720±100nM (n = 2) (Baaske et al, 2010) |
Optical Thermophoresis | 50% serum | Hill Equation | |
| Kd = 94.4 ± 26.6 nM (n = 1.13) (Nallagatla et al, 2009) |
Optical Thermophoresis | 20 mM Tris acetate pH 7.3, 140 mM NaCl, 5mM KCl, 1 mM MgCl2 and 1 mM CaCl2 | Single site binding curve from Origin 5 software package | |
| 20mer (modified 15mer) | Kd = 39±27nM (Hianik et al, 2007) |
Cyclic Voltametry | 140mM NaCl, 5mM KCl, 1mM CaCl2, 1mM MgCl2, 20mM Tris pH 7.4 | 1:1 from kinetics |
| 29mer | Kd = 0.5 nM (Tasset et al, 1997) |
Nitrocellulose Filter Binding | 50mM Tris-HCl, pH 7.5, 100mM NaCl, 1mM MgCl2 | Standard competitive binding model |
| Kd = 119.5nM (Lin et al, 2011) |
Surface Plasmon Resonance | 20mM Tris-HCl, 5mM KCl, 1mM MgCl2, 1mM CaCl2, 50mM NaCl, pH 7.4 | 1:1 from kinetics | |
| Kd = 31.1nM (Song et al, 2009) |
Capillary Electrophoresis with Laser Induced Fluorescence Polarization | 1xTG, pH 8.3, 5mM KCl | NECEEM | |
| Kd = 255±54nM (Li et al, 2008) |
Affinity Probe Capillary Electrophoresis | 2XTG, pH 8.5 | Simple isotherm | |
| Kd = 124.0±6.9 (n = 0.81) (Li et al, 2010) |
Affinity Probe Capillary Electrophoresis with Chemillumuniscent Detection | TGK 8.5 with HRP in anode vial TK with H2O2 in cathode vial TGKMg for sample prep | X, Y, double reciprocal plot | |
| 32mer (modified 29mer) | Kd = 88±53 (Hianik et al, 2007) |
Cyclic Voltametry | 140mM NaCl, 5mM KCl, 1mM CaCl2, 1mM MgCl2, 20mM Tris pH 7.4 | 1:1 reversible interaction from kinetics |