Automation of a Secondary Drug Screen for ALS Using iPSC-Derived Human Neurons


P Guyett1; M Martowicz2
1 BrainXell, Inc, United States;  2 Gilson Inc, United States


Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease caused by the death of motor neurons. A hallmark of ALS is low expression of neurofilament light chain (NFL) in motor neurons. A high-throughput phenotypic assay using patient-derived induced pluripotent stem cells (iPSCs) was used to screen over 6,000 compounds. Eighty compounds restored expression of NFL in reporter cell lines. These compounds were rescreened and validated by measuring reactivation of NFL expression in non-reporter lines by ELISAs. Given the error prone and time-consuming nature of secondary screening in 384-well plates, the process was automated on the Gilson PIPETMAX. The method prepared 8-point, 10-fold serial dilutions in triplicate across a 384-well plate followed by dosing of neurons. The automated screen was validated with a side-by-side comparison to manual pipetting. Two compounds, BX-1000 and BX-1200, were validated by the rescreening assay. Automation on PIPETMAX reduced hands-on time, eliminated human error, and had higher precision compared to manual pipetting.

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