Table 1.
Anti-STn mAb binding results summary.
| ELISA EC50 (nM) |
Glycan Array |
Internalization |
|||||||
|---|---|---|---|---|---|---|---|---|---|
| mAb | Isotype | BSM1 | OSM2 | PSM3 | (%)4 | Binding Group5 | FACS EC50 (nM)MDA-MB-231 STn+ | Mean FI | p-value |
| 8C2–2D6 | IgG2a,k | 0.4 | 0.5 | 0.3 | 99 | 1 | 2.8 | 7.6 | 0.03 |
| 2G12–2B2 | IgG2a,k | 0.1 | 0.1 | 0.1 | 99 | 1 | 0.3 | 6.7 | 0.01 |
| 4G8–1E3 | IgG2a,k | 1.2 | 0.6 | 0.1 | 99 | 1 | 0.7 | 7.4 | 0.01 |
| S3F* | IgG2a,k | 0.3 | 0.1 | 0.2 | 98 | 1 | 3.1 | 8.1 | 0.001 |
| 5G2–1B3 | IgG1,k | 0.4 | 0.2 | 10.2 | 92 | 4 | 0.2 | ND | ND |
| 2C2–2C5 | IgG3,k | 1 | 0.2 | 0.2 | 100 | 1 | 0.8 | 6.2 | 0.03 |
| 5E6–2E7 | IgG3,k | 5.8 | 0.8 | 0.4 | 100 | 1 | 1.8 | 7.5 | 0.04 |
| 9F11–1F7 | IgG3,k | 4.6 | 0.9 | 0.9 | 99 | 1 | 0.8 | 5.5 | 0.05 |
Summary Table of binding results.
1
BSM is bovine submaxillary mucin.
2
PSM is Porcine Submaxillary Mucin.
3
OSM is ovine submaxillary mucin.
4
% signal on the glycan array; the higher the percentage the more STn specific the antibody.
5
The binding group see Fig. 1. ND is not determined. ELISA EC50s were determined after subtracting signal from periodate treated wells from non-treated wells.
*
S3F was not generated during our immunizations – this is derived from HB-STn clone 3F1. We class switched from mouse IgG1 and recombinantly expressed as a mouse IgG2a, we refer to this engineered antibody as “S3F” in the text.