Table 1.
In vitro neutralizing activity by mAbs to HGF/SF
| mAb combination* | mAbs: Human HGF/SF (molar ratio)
|
|
|---|---|---|
| MDCK cells scattering† | Branching morphogenesis‡ | |
| Pool A (10 mAbs) | 80:1 | ND |
| Pool B (10 mAbs) | Neg. | ND |
| Pool C (11 mAbs) | Neg. | ND |
| A-1, -4, -5, -7, and -10 | 24:1 | 10:1 |
| A-1, -4, -5, and -7 | 20:1 | 10:1 |
| A-1, -4, -5, and -10 | 20:1 | 10:1 |
| A-1, -4, -7, and -10 | 20:1 | 10:1 |
| A-1, -5, -7, and -10 | 20:1 | 10:1 |
| A-1, -4, and -5 | Neg. | Neg. |
| A-1, -4, and -7 | 60:1 | 40:1 |
| A-1, -4, and -10 | 60:1 | 20:1 |
| A-1, -5, and -7 | 30:1 | 10:1 |
| A-1, -5, and -10 | 30:1 | 10:1 |
| A-1, -7, and -10 | 240:1 | Neg. |
| A-4, -5, and -7 | Neg. | 40:1 |
| A-4, -5, and -10 | Neg. | 20:1 |
| A-4, -7, and -10 | Neg. | 40:1 |
| A-5, -7, and -10 | Neg. | 40:1 |
ND, not done; Neg., negative.
mAb isotyping: A-1, -4, -5, and -10 are IgG1/κ; A-7 is IgG2b/κ.
*
mAbs A-2, -3, -6, -8, and -9 did not contribute to neutralization.
†
Scatter assay: all mAbs were adjusted to a final concentration of 2.0 mg/ml in PBS, 2.5 μl of each mAb pool were added to the first well, and 2-fold diluted to well 12. The end point is the mAb concentration that prevents scattering at the highest dilution. HGF/SF is 20 ng/ml, 250 μl per well.
‡
Branching morphogenesis assay; HGF/SF is 250 ng/ml, 200 μl per well. mAbs were 0.2, 0.4, 0.8, 1.6, and 3.2 μg per 200 μl; results were read and photographed after a 96-h incubation.