Table 4.
Summary of the bioassays performed in the collaborative study.
| Bioassay Type | Cell Line | No of Participants | TNF (IU/ml) | Assay Period (hrs) | Assay Readout | Readout Reagent |
|---|---|---|---|---|---|---|
| TNF Neutralisation Assays | ||||||
| Cytotoxicity | WEHI-164 | 9 | 2–100 | 18–24 | Absorbance | WST-8/CCK-8, MTS |
| L929 | 8 | 7.2–20 | 18–24 | Fluorescence, Absorbance, Luminescence | Resazurin/Alamar Blue, CCK-8, Cell-Titer Glo | |
| Apoptosis | U937 | 3 | 40–60 | 2.5 | Luminescence | Caspase Glo 3/7 |
| Rep-Gene | HEK 293 | 2 | 17.2–50.0 | 16–24 | Luminescence | Steady Glo |
| KLJ | 1 | 40 | 4 | Luminescence | Steady-Glo Plus | |
| Other Assays – target cell-lines express non-cleavable human TNF | ||||||
| CDC | Target: Jurkat | 2 | - | 4–22 | Absorbance, Luminescence | CCK-8, Cyto-Tox Glo |
| ADCC | Target: 3T3 cells, Effector: CD16 expressing NK92 | 1 | - | 4 | Luminescence | Cyto-Tox Glo |
| Target: CHO-K1 cells, Effector: CD16 expressing Jurkat cells linked to luciferase reporter gene | 1 | - | 16–24 | Luminescence | Bio-Glo | |
| Target: HEK-293 cells, Effector: CD16 expressing Jurkat cells linked to luciferase reporter gene | 1 | - | 6 | Luminescence | Dual Glo | |