Confirm true
binders in
relevant biology
Hit confirmation focuses on verifying that initial screening hits represent true, reproducible modulators of the target of interest. During this phase, preliminary hits are converted into decision-grade actives through systematic retesting and careful assay design.
Choices regarding cell model and assay format are made to ensure biological relevance and accuracy. At the same time, control design, plate layout, and quality control metrics are defined to support the generation of robust and reproducible data. Retesting strategies typically include single or multiple concentration measurements and concentration–response experiments to estimate potency and consistency of effect.
Hit confirmation also establishes the criteria for compound progression into follow-up biological studies and entry into Structure–Activity Relationship analysis. Counter-screens and interference checks are incorporated to rule out nonspecific effects, while orthogonal evidence is generated to strengthen confidence in on-target activity. Together, these decisions ensure that only well-characterized compounds with clear cellular target engagement advance to the next stage of discovery.
Confirm hits with confidence: Targeted Profiling
CETSA applied in relevant cellular models provides an additional layer of confidence by verifying direct target engagement under retesting conditions. This helps separate genuine actives from assay-dependent effects and supports clear progression criteria for compounds moving into SAR-driven optimization.
Hit confirmation is enabled by our Targeted Profiling Service
Targeted Profiling
Validate mechanistic hypotheses and confirm that on-target engagement occurs in the intended cellular context.
Is your hit on target?
Learn how you can identify, validate, and optimize hits directly in unmodified cellular systems.
Are you using the right tools?
Early integration of CETSA technology can help you streamline your discovery pipeline, minimize late-stage failures, and uncover novel targets faster than traditional methods.
What is CETSA
Why work directly in cells?
Why Drug Candidates Fail
Why is Target ID so important?
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