hit-to-lead discovery

UbiQ can assist your lead discovery in the UPS with our wide range of activity assay reagents. Perfect tools for high throughput compound screening;

UbiQ Assay

UbiQ-Assay, DUB activity assay reagents, proved effective in multiple industrial scale HTS campaigns. They are available in various flavors for a wide range of assay read outs, e.g. fluorogenic substrates for fluorescence intensity read-out, substrates for FP assays, or as highly specialized context specific FP reagents (our class II FP reagents). All of them targeting DUBs & Ub-like proteases.

 

applications

  • compound library screening
  • activity assays

 

targets

  • DUBs and Ubl proteases

 

experience

  • substrate supply for an undisclosed number of large library screens by our industrial partners
  • development of >10 highly specialized context specific custom 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’-valuevalidated hits

hit selected for

dose-response

DUB-001UbiQ-012 (Ub FP)UbiQ-002 (Ub-Rh110)225K0.7 – 0.8293 (0.14%)99 (0.05%)
310K0.7 – 0.8664 (0.21%)182 (0.06%)
DUB-002UbiQ-012 (Ub FP)UbiQ-002 (Ub-Rh110)310K0.7 – 0.8516 (0.17%)152 (0.05%)
DUB-003Ub-like FP reagentUb-like (AMC)310K0.7 – 0.81009 (0.33%)170 (0.05%)
DUB-004Ub-like FPUb-like (AMC)310K0.7 – 0.8ongoingongoing
DUB-005Ub FP class II reagentUbiQ-002 (Ub-Rh110)310K0.75 – 0.8ongoingongoing

* 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