The design, fabrication, and preliminary testing is presented for a polymer multilayered hybrid micro-nanofluidic chip that consists of poly(methylmethacrylate) (PMMA) layers containing microfluidic channels separated in the vertical direction by polycarbonate (PC) nanocapillary array membranes (NCAMs). This design architecture enables nanofluidic interconnections to be placed in the vertical direction between microfluidic channels. Such an architecture combines microfluidic manipulations (separation, injection, collection, etc.) with nanofluidic molecular capabilities (molecular sizing and affinity reactions, channel isolation, enhanced mixing, etc.) on a single chip. Recent polymeric microfabrication advances have made this scalable construct possible: 1) processing thin polymer layers on releasable and compliant carriers, and 2) the high resolution contact-printing of a strong thermal adhesive. Bond strength was demonstrated by pressurizing channels with 90 psi nitrogen without failure. Devices were characterized in terms of measuring resistivity and electroosmotic flow (EOF) along the channels at different pH values. The functionality of the chip is demonstrated by filling a cross channel with 1 µM green-fluorescent protein (GFP) and electrokinetically transporting analyte plugs through the NCAM and down the separation channel while performing laser induced fluorescence (LIF) analysis. The development of this new type of hybrid micro-nanofluidic device potentially will allow unprecedented molecular manipulations for chemical and biological sensing applications.