Table 1.
Clone characteristics
| scFv phage binder |
scFv SNAP binder |
scFv Fc binder |
Affinity |
Internalization |
||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Clone | incidence | ELISA | FACS | ELISA | FACS | FACS | IHC | KD ± SD in nM | FACS | Confocal |
| EMI404 | 1x | ++ | ++ | ++ | ++ | + | n.d. | 62.9 ± 6.8 | ++ | + |
| EMI405 | 8x | + | ++ | + | ++ | ++ | ++ | 63.9 ± 8.3 | ++ | ++ |
| EMI406 | 13x | + | ++ | + | ++ | - | n.d. | 159.4 ± 87.5 | ++ | ++ |
| EMI407 | 3x | + | + | + | ++ | - | n.d. | 404.4 ± 183.0 | + | ++ |
Selected binders are categorized as moderate (+) or strong (++) based on the ELISA absorption value (+ < 5x, ++ > 5x higher than background), the percentage of shifted cells identified by FACS (+ < 60%, ++ > 60%), the fluorescence intensity under confocal microscopy (+ high, ++ very high) and the immunohistochemical (IHC) staining intensity (+ moderate, ++ strong). n.d. = not determined. Experiments were carried out at least three times.