A fast and reliable method for the detection of molecular interactions in real‐time is necessary for drug screening and control of infections or diseases. Therefore, this study aims to develop a high‐throughput portable nanoplasmonic platform. This is done by integrating a nanocup array‐enhanced surface plasmon resonance (NanoSPR) sensor with a standard 96‐well plate or a simple eight‐pillar device, which can offer rapid and label‐free interaction analysis and highly sensitive binding kinetics for drug screening. To fabricate the wafer‐level NanoSPR biosensor, nanoimprint lithography, electron beam evaporation, and bonding technology have been used to offer low‐cost production on a mass scale. Meanwhile, a variety of ready‐to‐use NanoSPR biosensors are separately modified to demonstrate the high‐quality binding kinetics and affinity of different biomolecular interactions. Owing to their unique optical properties, the two developed portable NanoSPR devices provide the possibility of having a portable benchtop molecular interaction instrument that is very promising for meeting the needs of personal and laboratory testing. This study may pave the way for NanoSPR biosensors to achieve high‐throughput and easy‐to‐use molecular interaction analysis with outstanding performance. [ABSTRACT FROM AUTHOR]