High-Content Imaging for Large-Scale Detection of Low-Affinity Extracellular Protein Interactions.

ABSTRACT: Extracellular protein interactions coordinate cellular responses with their local environment and have important roles in pathogen invasion and disease. Due to technical challenges associated with studying binding events at the cell surface, the systematic and reliable identification of novel ligand-receptor pairs remains difficult. Here, we describe the development of a cell-based assay using large-scale transient transfections and high-content imaging (HCI) to detect extracellular binding events. We optimized the parameters for efficient transfection of human cells with cDNA plasmids encoding full-length cell surface receptors in 384-well plates. Using a range of well-characterized structurally diverse low-affinity cell surface interactions, we show that transfected cells probed with hig

SUBMITTER: Wood L 

PROVIDER: S-EPMC6873222 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature