Formation of Tethered Bilayer Lipid Membranes (tBLMs) Using Plasma Membrane Blebs from Human Lung Cancer Cells

Abstract Plasma membrane blebs, or giant plasma membrane vesicles (GPMVs), serve as excellent biomimetics for studying the compositional complexity of native cell membranes. However, current supported membrane models often lack the stability or submembrane reservoir necessary for extended functional studies. This work presents a proof of concept for the formation of tethered bilayer lipid membranes (tBLMs) using blebs derived from human lung cancer cells (A549). We report that successful fusion of blebs into planar tBLMs on gold substrates requires a significantly higher density of tethering anchors (60 mol % WC14) than that required for synthetic liposomes. Surface-enhanced infrared absorption spectroscopy (SEIRAS) analysis indicates that this high anchor density promotes a vertical alignment of the alkyl chains, thereby facilitating the fusion of the complex native membranes. To demonstrate the utility of this platform for pharmacological screening, we investigated the membrane-disrupting mechanism of the anticancer drug edelfosine. Fluorescence lifetime imaging microscopy (FLIM) revealed that edelfosine reduces membrane microviscosity, consistent with a detergent-like permeabilization mechanism. These findings establish bleb-derived tBLMs as a robust platform for applying surface-sensitive analytical techniques to the study of native membrane properties and drug–membrane interactions.