Cellular Assays with a Molecular Endpoint Measured by SAMDI Mass Spectrometry.

ABSTRACT: Cell-based, high-throughput screening (HTS) assays are increasingly important tools used in drug discovery, but frequently rely on readouts of gene expression or phenotypic changes and require development of specialized, labeled reporters. Here a cell-based, label-free assay compatible with HTS is introduced that can report quantitatively on enzyme activities by measuring mass changes of substrates with matrix-assisted laser desorption/ionization mass spectrometry. The assay uses self-assembled monolayers to culture cells on arrays presenting substrates, which serve as reporters for a desired enzyme activity. Each spot of cells is treated with a compound, cultured and lysed, enabling endogenous enzymes to act on the immobilized peptide substrate. It is demonstrated that the assay can measu

SUBMITTER: Berns EJ 

PROVIDER: S-EPMC4981186 | biostudies-literature | 2016 Jul

REPOSITORIES: biostudies-literature