Application Notes

Ultra-high-throughput biophysical screening of MEK1 using the Dianthus uHTS platform

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8 A P P L I C A T I O N N O T E Equation. 2 Automated Screening Platform Dianthus uHTS was integrated with a ThermoFisher Orbiter RS1 arm, and workflow control was managed via Momentum Scheduler (v. 6.1). Target dispense was performed with a continuous flow chip on a Formulatrix Mantis (v. 4.5.1), shaking on a BioShake 5000 elm (QInstruments GmbH), barcodes scanned using a SR-1000 (Keyence Germany GmbH), centrifuged with a VSpin (Agilent) and sealed with a PlateLoc (Agilent). Plates were incubated at 25 °C in a Cytomat (ThemoFisher). Dianthus uHTS was controlled via gRPC and internally built python-based driver. ANALYSIS OF DATA Data analysis was performed in a custom built pipeline in Streamlit (v1.40.0) [15], using Python 3.11.9 [16] with the following packages: altair (v5.5.0), click (v8.1.7), dantte (v0.0.0.dev570+dianthusuhts, an internal NanoTemper Robust Fitting Library for well scans and dose response curves), GitPython (v3.1.44), matplotlib (v3.7.2), numpy (v1.24.3), pandas (v2.0.3), plotly (v5.9.0), pytest (v8.3.2), PyYAML (v6.0.2), scikit_learn (v1.6.1), scipy (v1.15.2), seaborn (v0.13.2), soupsieve (v2.6), statsmodels (v0.14.2), streamlit (v1.40.1), streamlit_extras (v0.6.0), streamlit_option_menu (v0.4.0), tinydb (v4.8.2), typer (v0.15.2). Assay quality was monitored by calculated Robust Z' values for each plate using DMSO reference as negative control and U0126 as positive control. Equation 3 is used to calculate the robust Z' value, where median (x̃ ) and standard deviation equivalent (σ robust ) of the Median Absolute Deviation (MAD, Equation 5). The MAD (Equation 4) can be easily calculated using standard statistical packages. Equation. 3

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