In this project, a functional biomimetic membrane utilizing the highly permeable and mutation-tolerant E. coli Outer Membrane Protein F (OmpF) was developed. OmpF was self-assembled with the block copolymer polybutadiene-polyethylene oxide (PB-PEO) to form two-dimensional crystals using a dialysis technique. The E. coli OmpF was overexpressed in an expression strain and purified through a series of centrifugations and chromatographic steps. Once the OmpF was crystallized with the PB-PEO, transmission electron microscopy imaging was utilized to analyze morphology of the single layer. Suitable single layers were then stacked in a layer-by-layer method to produce a functional membrane. This membrane was subsequently characterized to obtain its water permeability and solute rejection data for dyes of various molecular weights and sizes. This experiment indicated that OmpF can be successfully incorporated into a PB-PEO membrane and can be used for separations of molecules with molecular weights greater than 500 Da with high permeability.