(182c) Recombinant ELP Bolaamphiphiles for the Controlled Construction of Globular Protein Vesicles

Synthetic biology is rapidly advancing the design of artificial cells capable of sensing and responding to their environment. This work presents a modular approach to engineering globular protein vesicles (GPVs) using fully recombinant components. Specifically, we developed multifunctional elastin-like peptide (ELP) bolaamphiphiles that self-assemble into vesicle membranes and display folded proteins at both termini. Using a streamlined Golden Gate-based cloning strategy, we efficiently designed, expressed, and purified a library of ELP bolaamphiphiles with varying structural complexity, including fluorescent protein domains, for visualization and characterization. By tuning the molar ratios of these building blocks, we optimized the self-assembly and morphology of GPVs. Characterization by fluorescence imaging and dynamic light scattering confirmed vesicle formation and revealed size distributions influenced by bolaamphiphile design. These results highlight the potential of ELP-based bolaamphiphiles for modular GPV construction, offering a platform for building synthetic cells with customizable functionalities and controllable architectures.