Table 1.
Example determination of F:P ratios for ALK-APC and L1CAM-BV421 antibody conjugates
| MFI values |
||||||
|---|---|---|---|---|---|---|
| Labeled antibody | BV421 | PE | APC | Fluorophore molecules/cell | F:P ratio | Antigen molecules/cell |
| ALK-APC | 595 | 4325 | 3.34b | 1296 | ||
| ALK-PE | 269 | 1296 | 1:1a | 1296 | ||
| L1CAM-BV421 | 864 | 8041 | 2.21b | 3632 | ||
| L1CAM-PE | 788 | 3632 | 1:1a | 3632 | ||
Determination of fluorophore-to-protein ratios of a given monoclonal antibody (ALK-APC and L1CAM-BV421) by staining neuroblastoma cells with these mABs and the same clone labeled in PE with 1:1 F:P ratios, respectively. Quantibrite-PE and quantitation beads in APC or BV421 allow calculation of F:P from fluorophore molecules/cell with equivalent antigen molecules/cell.
a
Known F:P ratio.
b
Calculated F:P ratio as detailed in Table 3 and according manuscript section.