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Table 3 Kinetic parameters obtained from the optimization of the Peptidomatrix conditions with the peptides and antibodies specific to Na/K ATPase (see figure 4) (± SD, n = 3)
| A: increasing attached peptide at two fixed levels of added antibody | |||
|---|---|---|---|
| 1 stAB Concentration 0.00033 serum/PBS | 1 stAB Concentration 0.00066 serum/PBS | ||
| Kd (μg/ml) | 0.19 ± 0.01 | 0.1 ± 0.02 | |
| Amax (OD units) | 1.36 ± 0.02 | 1.56 ± 0.08 | |
| D (OD units) | 0.25 ± 0.02 | 0.38 ± 0.07 | |
| B: Competition assay – Kd determined when increasing added free peptide at three fixed levels of added antibody | |||
| 1 stAB concentration 0.000625 serum/PBS | 1 stAB concentration 0.000125serum/PBS | 1 stAB concentration 0.0025 serum/PBS | |
| Kd (μg/ml) | 0.55 ± 0.1 | 0.83 ± 0.12 | 2.55 ± 1.21 |
| Amax (OD units) | 1.16 ± 0.05 | 1.35 ± 0.05 | 1.33 ± 0.17 |
| D (OD units) | 0.59 ± 0.04 | 0.86 ± 0.04 | 1.17 ± 0.18 |
| C: increasing attached peptide at two fixed levels of added antibody | |||
| 1 stAB concentration 0.00033 serum/PBS | 1 stAB concentration 0.00067 serum/PBS | ||
| Kd (serum dilutions) | 0.33 ± 0.08 | 0.29 ± 0.04 | |
| Amax (OD units) | 1.46 ± 0.09 | 1.74 ± 0.06 | |
| D (OD units) | 0.19 ± 0.079 | 0.24 ± 0.05 | |
| D: Competition assay- Kd determined when increasing added free peptide at three fixed levels of added antibody | |||
| 1 stAB concentration 0.000625 serum/PBS | 1 stAB concentration 0.00125 serum/PBS | 1 stAB concentration 0.0025 serum/PBS | |
| Kd (μg/ml) | 0.68 ± 0.12 | 0.99 ± 0.19 | 2.04 ± 0.3 |
| Amax (OD units) | 1.58 ± 0.06 | 1.87 ± 0.09 | 2.05 ± 0.08 |
| D (OD units) | 0.25 ± 0.05 | 0.25 ± 0.08 | 0.28 ± 0.08 |
