High-throughput detection of metal contamination in HTS outputs

Molyneux, Corinne; Sinclair, Ian; Lightfoot, Helen L.; Walsh, Jarrod; Holdgate, Geoffrey A.; Moore, Rachel

doi:10.1016/j.slasd.2022.03.003

Cited by 4 publications

(1 citation statement)

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“…Metal impurities leading to false positives in HTS campaigns have been described before. − We intended to use our learnings to comprehensively identify the most robust assay formats with the goal of reducing the likelihood of false-positive results. We conducted a systematic comparison of different biochemical KRAS screening configurations (Table ).…”

Section: Resultsmentioning

confidence: 99%

Chasing Red Herrings: Palladium Metal Salt Impurities Feigning KRAS Activity in Biochemical Assays

Gerstberger,

Berger,

Büttner

et al. 2024

J. Med. Chem.

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Identifying promising chemical starting points for small molecule inhibitors of active, GTP-loaded KRAS "on" remains of great importance to clinical oncology and represents a significant challenge in medicinal chemistry. Here, we describe broadly applicable learnings from a KRAS hit finding campaign: While we initially identified KRAS inhibitors in a biochemical high-throughput screen, we later discovered that compound potencies were all but assay artifacts linked to metal salts interfering with KRAS AlphaScreen assay technology. The source of the apparent biochemical KRAS inhibition was ultimately traced to unavoidable palladium impurities from chemical synthesis. This discovery led to the development of a Metal Ion Interference Set (MIIS) for up-front assay development and testing. Profiling of the MIIS across 74 assays revealed a reduced interference liability of label-free biophysical assays and, as a result, provided general estimates for luminescence-and fluorescence-based assay susceptibility to metal salt interference.

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Section: Resultsmentioning

confidence: 99%