hit-to-lead discovery
UbiQ can assist your lead discovery in the UPS with our wide range of activity assay reagents. Perfect tools for your high-throughput screening campaign.
Our DUB activity assay reagents proved effective in multiple industrial scale high-throughput screening campaigns. They are available in different assay read-outs (e.g. fluorogenic, fluorescence polarization or bioluminiscence) and complexity (e.g. our class II FP reagents).
applications
- high-throughput screening
- activity assays
targets
- DUBs and Ubl proteases
experience
- substrate supply for an undisclosed number of large library screens by our industrial partners
- custom development of more than 10 highly specialised context specific assay reagents
- demonstrated performance at independent European screening centers – Pivot Park Screening Center and European ScreeningPort (table 1)
- five DUB screening campaigns (>1.7 million small molecules)
table 1: UbiQ’s screening reagents used in HTS campaigns*
| screen# | primary screen + dose-response | orthogonal assay for hit validation | small-molecule library size | Z’-value | validated hits | hit selected for dose-response |
| DUB-001 | UbiQ-012 (Ub FP) | UbiQ-002 (Ub-Rh110) | 225K | 0.7 – 0.8 | 293 (0.14%) | 99 (0.05%) |
| 310K | 0.7 – 0.8 | 664 (0.21%) | 182 (0.06%) | |||
| DUB-002 | UbiQ-012 (Ub FP) | UbiQ-002 (Ub-Rh110) | 310K | 0.7 – 0.8 | 516 (0.17%) | 152 (0.05%) |
| DUB-003 | Ub-like FP reagent | Ub-like (AMC) | 310K | 0.7 – 0.8 | 1009 (0.33%) | 170 (0.05%) |
| DUB-004 | Ub-like FP | Ub-like (AMC) | 310K | 0.7 – 0.8 | ongoing | ongoing |
| DUB-005 | Ub FP class II reagent | UbiQ-002 (Ub-Rh110) | 310K | 0.75 – 0.8 | ongoing | ongoing |
* Completed and ongoing HTS programs in which UbiQ’s FP assay reagents were used for the primary screen (single point at 10 μM), and dose-response experiments of validated hits. All assays showed excellent Z’ values (>0.7). After confirmation of the hits with the FP substrate, an orthogonal Ub-Rh110Gly (UbiQ-002) or Ub-like AMC assay was used for validation; the number of hits identified at this stage varied from 0.33−0.14% of the total library. A subset of compounds (in general 0.05% of the total library) is then selected for IC50 determination using the FP assay reagent (10 concentration steps from 0.15 nM−5 μM).
> See UbiQ-Assay fact sheets for more information, available options and references. Also available in Japanese.