© 2016 IOP Publishing Ltd. Drug discovery is a complex and time consuming process involving significant basic research and preclinical evaluation prior to testing in patients. Preclinical studies rely extensively on animal models which often fail in human trials. Biomimetic microphysiological systems (MPS) using human cells can be a promising alternative to animal models; where critical interactions between different organ systems are recreated to provide physiologically relevant in vitro human models. Central here are blood-vessel networks, the interface controlling transport of cellular and biomolecular components between the circulating fluid and underlying tissue. Here we present a novel lab-on-a-brane (or lab-on-a-membrane) nanofluidics MPS that combines the elegance of lab-on-a-chip with the more realistic morphology of 3D fibrous tissue-engineering constructs. Our blood-vessel lab-on-a-brane effectively simulates in vivo vessel-tissue interface for evaluating transendothelial transport in various pharmacokinetic and nanomedicine applications. Attributes of our platform include (a) nanoporous barrier interface enabling transmembrane molecular transport, (b) transformation of substrate into nanofibrous 3D tissue matrix, (c) invertible-sandwich architecture, and (d) simple co-culture mechanism for endothelial and smooth muscle layers to accurately mimic arterial anatomy. Structural, mechanical, and transport characterization using scanning electron microscopy, stress/strain analysis, infrared spectroscopy, immunofluorescence, and FITC-Dextran hydraulic permeability confirm viability of this in vitro system. Thus, our lab-on-a-brane provides an effective and efficient, yet considerably inexpensive, physiologically relevant alternative for pharmacokinetic evaluation; possibly reducing animals used in preclinical testing, costs from false starts, and time-to-market. Furthermore, it can be configured in multiple simultaneous arrays for personalized and precision medicine applications and for evaluating localized and targeted therapeutic delivery.