Assessing Trehalose-Mediated Stabilization of Liposomes using Calcium Permeability Assay
Liposome-based systems serve as essential membrane-bound compartments in synthetic cells however, their long-term stability is often limited by membrane leakage. This project aims to determine whether the disaccharide sugar trehalose can reduce liposome permeability and improve membrane stability under temperature-induced stress including multiple freeze-thaw cycles. To test this, liposomes containing calcium chloride will be formed and transferred to an exogenous buffer containing the calcium-sensitive fluorescent indicator Indo-1 pentapotassium salt (IPPS). IPPS is unable to freely cross the lipid bilayer, therefore, changes in fluorescence will directly indicate calcium leakage from the liposome lumen into the surrounding solution. I hypothesize that trehalose will reduce calcium leakage and preserve liposome integrity during thermal stress. This study may support the development of more durable liposome-based systems for synthetic biology and enhance the stability of long-term storage applications.