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).
- high-throughput screening
- activity assays
- DUBs and Ubl proteases
- 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*
primary screen +
assay for hit validation
hit selected for
|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).