Determining the expression level of biomarkers is crucial for disease diagnosis. However, the low abundance of biomarkers in the early stage makes the detection extremely difficult by traditional aggregation-induced emission (AIE)-based fluorescent probes. Here, by tuning the intermolecular interaction, a two steps-based MP/NPs-SLIPS sensing system is designed for ultrasensitive detection of the tumor marker matrix metalloproteinase-2 (MMP-2). During the sensing process, aggregation of AIE residual could be intensified through the electrostatic absorption by negatively charged nanoparticles (NPs), as well as the confined space formed by the self-assembly of NPs to photonic crystals (PCs) on slippery lubricant-infused porous substrates (SLIPS). The fluorescent signals obviously increased with a strengthened aggregation degree, which contributes to improved sensitivity. Thus, the limit of detection is decreased to 3.7 ng/mL for MP/NPs-SLIPS sensing system, which could be used for detecting the MMP-2 secreted by tumor cells directly. This strategy also demonstrated its potential applications as high-throughput detection devices and will be of significance for the ultrasensitive analysis of biomarkers.