One-Pot Time-Induced\nProteome Integral Solubility\nAlteration Assay for Automated and Sensitive Drug–Target Identification

The proteome integral solubility alteration (PISA) assay\nis widely\nused for identifying drug targets, but it is labor-intensive and time-consuming\nand requires a substantial amount of biological sample. Aiming at\nenabling automation and greatly reducing the sample amount, we developed\none-pot time-induced (OPTI)-PISA. Here, we demonstrate OPTI-PISA performance\non identifying targets of multiple drugs in cell lysate and scaling\ndown the sample amount to sub-microgram levels, making the PISA method\nsuitable for NanoProteomics. OPTI-PISA can be implemented using only\nthe standard equipment of a proteomics lab.