Biomarker Nanoparticles for Non-Invasive Enzyme Profiling

The patent uses an intermediary approach by administering protease-sensitive nanoparticles as a mediator that indirectly detects protease activity. Instead of directly sampling tissue or blood through invasive procedures, the nanoparticles circulate systemically, are selectively cleaved by target proteases at disease sites, and release detectable markers that are then non-invasively detected in urine samples. This intermediary detection mechanism maintains high reliability while eliminating the need for invasive sampling procedures.

The patent employs a copying strategy by designing detectable markers that are released upon protease cleavage of the nanoparticles. These markers are engineered to be highly specific and can be detected using simplified affinity-based assays that replicate the precision of mass spectrometry without requiring the complex instrumentation. The markers serve as simplified copies or proxies for the original complex biomarker detection approach, maintaining measurement precision while dramatically reducing device complexity and cost.

The patent applies segmentation by dividing the detection system into multiple functional components: (1) protease-sensitive nanoparticles that serve as circulating sensors, (2) enzyme-cleavable linkers that act as molecular switches, and (3) detectable markers that provide the signal. This segmented architecture allows each component to be optimized independently, enabling ultra-sensitive detection of early-stage disease where protease activity is just beginning to occur. The segmentation also enables multiplexing to detect multiple proteases simultaneously, further improving diagnostic accuracy.