Gram-negative bacteria are enclosed by an asymmetric outer membrane that protects against antibiotics and antimicrobial peptides by serving as an impermeable barrier. With the rise in antibiotic resistance, in-vitro tools such as bacterial membrane models have become critical for understanding the intrinsic properties of Gram-negative outer membrane that impact membrane permeability and their effects on antibiotic efficacy. However, these membrane models often lack the molecular and structural complexity of the native Gram-negative outer membrane. To overcome this, we developed a Gram-negative outer membrane model that captures key components of the outer membrane as a platform for studying membrane behavior in presence of antibiotics. This membrane model utilizes outer membrane vesicles (OMVs) derived from the outer membrane of clinically relevant Gram-negative pathogens to incorporate native membrane material. For this dissertation, I demonstrated the formation of supported bilayer using OMVs from various Gram-negative isolates on glass and on conductive surfaces, and the retention of membrane mobility and native membrane components in our system. In Chapter 2 and 3, I illustrated the use of outer membrane models in studying antibiotic-membrane interactions using polymyxin B and bacitracin with tools such as quartz crystal microbalance with dissipation (QCM-D) and electrochemical impedance spectroscopy (EIS). In Chapter 4 and 5, I further expanded this platform to study outer membrane biomolecular interactions involved in indirect electron transfer in the presence or absence of quantum dots and measuring antibiotic accumulation through use of boron-detecting sensors. With this system, we demonstrate a robust and adaptable platform that can be used to investigate biomolecular properties of outer membrane from any Gram-negative isolate while retaining native components and antibiotic-membrane behavior.