Molecular binding kinetics quantification is pivotal in the design of antibodies, small-molecule drugs, and early diagnosis of complex disease. This study presents a portable microfluidic system, featuring a nanoplasmonic sensor with cavity reflection enhancement (NSCRE), designed for the ultrasensitive detection and analysis of molecular interactions. This device comprises liquid flow paths, generic fiber spectrometers, a versatile NSCRE sensor chip, and a novel data analysis system. Furthermore, a neural network data processing system is developed based on a deep learning model to decrease significant data fluctuations caused by air segments. This “all-in-one” platform, integrating multifunctional, easy-to-use, and renewable NSCRE biosensors, boasts detection capabilities for both slow and rapid kinetics. It can automatically monitor interactions between molecules across a broad range of molecular weights (0.259–150 kDa), allowing for real-time, ultra-sensitive, and highly selective analysis. In addition, the platform with CRE effect facilitates a 7.2-fold increase in reflection intensity signals compared to that of a sensor without CRE. The immunoglobulin G detection with a limit of detection of 73 pM based on this device, showing a 50-fold increase in sensitivity compared to previously reported values. The miniaturized automatic detection system paves a new path for the use of molecular interaction analysis in life science research, drug discovery, early diagnosis, and other fields.
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