Table 3.
Inhibition of IgE binding as measured by microarray and by ImmunoCAP after addition of blocking IgG. Different concentrations of monoclonal human Bet v 1-specific IgE (chim.IgE; concentrations indicated in coloured boxes) were incubated with increasing concentrations of blocking IgG (relative excess compared to IgE and, in parentheses, absolute concentrations of IgG indicated in the left columns) and measured by microarray (left tables) or ImmunoCAP (right tables). Absolute responses after subtraction of background signals are indicated in ISU for the MeDALL allergen-chip and in UA/ml for ImmunoCAP. Relative inhibition was calculated for every IgE-concentration applied (baseline responses indicated in red numbers).
| MeDALL-chip | ||||
|---|---|---|---|---|
| chim. IgE → | 312.5 ng/ml |
|||
| Absolute response | % Inhibition | |||
| Blocking IgG ↓ | Mean | SD | Mean | SD |
| 0× (0 ng/ml) | 92.86 | 2.72 | 0.00 | 2.92 |
|
| ||||
| 1× (312.5 ng/ml) | 52.15 | 6.13 | 43.84 | 6.60 |
|
| ||||
| 10× (3.1 μg/ml) | 20.05 | 1.39 | 78.41 | 1.50 |
|
| ||||
| 100× (31.2 μg/ml) | 2.60 | 0.42 | 97.20 | 0.46 |
|
| ||||
| 1000× (312.5 μg/ml) | 0.32 | 0.02 | 99.66 | 0.02 |
| chim. IgE → | 6.25 ng/ml |
|||
|---|---|---|---|---|
| Absolute response | % Inhibition | |||
| Blocking IgG ↓ | Mean | SD | Mean | SD |
| 0× (0 ng/ml) | 5.58 | 0.62 | 0.00 | 11.17 |
|
| ||||
| 1× (6.25 ng/ml) | 5.04 | 0.46 | 9.56 | 8.28 |
|
| ||||
| 10× (62.5 ng/ml) | 4.56 | 0.74 | 18.23 | 13.27 |
|
| ||||
| 100× (625 ng/ml) | 1.62 | 0.20 | 70.95 | 3.52 |
|
| ||||
| 1000× (6.25 μg/ml) | 0.18 | 0.03 | 96.83 | 0.62 |
|
| ||||
| 10,000× (62.5 μg/ml) | 0.02 | 0.02 | 99.58 | 0.27 |
| chim. IgE → | 0.8 ng/ml |
|||
|---|---|---|---|---|
| Absolute response | % Inhibition | |||
| Blocking IgG ↓ | Mean | SD | Mean | SD |
| 0× (0 ng/ml) | 0.65 | 0.10 | 0.00 | 16.14 |
|
| ||||
| 1× (0.8 ng/ml) | 0.65 | 0.06 | 0.00 | 8.61 |
|
| ||||
| 10× (8 ng/ml) | 0.68 | 0.13 | −4.64 | 20.10 |
|
| ||||
| 100× (80 ng/ml) | 0.46 | 0.14 | 28.35 | 22.37 |
|
| ||||
| 1000× (800 ng/ml) | 0.09 | 0.04 | 85.57 | 6.25 |
|
| ||||
| 10,000× (8 μg/ml) | 0.03 | 0.00 | 95.88 | 0.00 |
|
| ||||
| 100,000× (80 μg/ml) | 0.00 | 0.01 | 100.00 | 0.89 |
| Bet v 1 ImmunoCAP | ||||
|---|---|---|---|---|
| chim. IgE → | 312.5 ng/ml |
|||
| Absolute response | % Inhibition | |||
| Blocking IgG ↓ | Mean | SD | Mean | SD |
| 0× (0 ng/ml) | 91.11 | 3.35 | 0.00 | 3.67 |
|
| ||||
| 1× (312.5 ng/ml) | 83.14 | 2.30 | 8.74 | 2.53 |
|
| ||||
| 10× (3.1 μg/ml) | 90.91 | 3.95 | 0.22 | 4.34 |
|
| ||||
| 100× (31.2 μg/ml) | 91.74 | 5.41 | −0.70 | 5.94 |
|
| ||||
| 1000× (312.5 μg/ml) | 21.54 | 1.16 | 76.36 | 1.27 |
| chim. IgE → | 20 ng/ml |
|||
|---|---|---|---|---|
| Absolute response | % Inhibition | |||
| Blocking IgG ↓ | Mean | SD | Mean | SD |
| 0× (0 ng/ml) | 5.39 | 0.26 | 0.00 | 4.76 |
|
| ||||
| 1× (20 ng/ml) | 5.59 | 0.10 | −3.71 | 1.90 |
|
| ||||
| 10× (200 ng/ml) | 5.84 | 0.06 | −8.34 | 1.02 |
|
| ||||
| 100× (2 μg/ml) | 6.05 | 0.20 | −12.11 | 3.75 |
|
| ||||
| 1000× (20 μg/ml) | 6.17 | 0.08 | −14.34 | 1.52 |
|
| ||||
| 10,000× (200 μg/ml) | 2.29 | 0.26 | 57.48 | 4.83 |