[PEER REVIEWED] Cell surface Liposome Binding (CLiB) assay v1

This protocol describes a workflow for the Cell surface Liposome Binding (CLiB) assay to analyze protein–lipid interactions. The procedure covers buffer preparation, construction of yeast display libraries, liposome preparation, flow cytometry–based analysis, and next-generation sequencing (NGS) analysis. Using this workflow, it is possible to rapidly identify and engineer lipid-binding domains with defined specificity and affinity for diverse membrane lipids. The protocol can be applied both to the analysis of lipid-binding domains and to on-demand generation of lipid-binding probes.