MOLECULARLY-COMPLETE PLANAR SUPPORTED CELL PLASMA MEMBRANES AS SCAFFOLDS FOR BIOMEDICAL APPLICATIONS
Emerging technologies to study membrane proteins, protein-lipid interactions, and to create new kinds of sensing and analytical devices, use cell plasma membrane vesicles or ‘blebs’ as an intermediate to form molecularly complete, planar cell surface mimetics that are compatible with a variety of characterization tools and microscopy methods. This approach enables direct incorporation of membrane proteins into supported lipid bilayers without using detergents and reconstitution, preserving the native lipids and other species within the plasma membrane. However, the impact of methods used to induce cell blebbing (vesiculation) on protein and membrane properties is still unknown. This study focuses on characterization of the cell blebs created under various bleb-inducing conditions and its result on protein behavior (orientation, mobility, activity, etc.) and lipid scrambling in this platform. This work enriches our understanding of cell plasma membrane bleb bilayers as a biomimetic platform and represents one of few ways to make molecularly-complete supported bilayers from cell membranes. Such a model system can be widely applied to studies aimed at understanding the roles of membrane proteins as drug targets in drug delivery, in virus-host interactions, and in tissue engineering platforms, among many other bioanalytical and sensing applications. Here, we applied this biomimetic model system to studies of oncogenic micrcrovesicle interaction with stem cell surface. Combined with TIRF, the impact of microenvironment on binding and entry of microvesicle to stem cell surface will be revealed at the single particle level.
Biomimetic platform; Cell plasma membrane vesicles; Chemical engineering; Single particle tracking; Supported lipid bilayer; Protein diffusion
Paszek, Matthew J.; Fischbach, Claudia
M.S., Chemical Engineering
Master of Science
dissertation or thesis