Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) is typically diagnosed at an advanced stage, when curative options are limited. Circulating extracellular vesicles (EVs) provide a minimally invasive window into tumor biology, yet existing EV assays are slow, sample-intensive, and incompatible with clinical workflows requiring speed, reproducibility, and minimal sample input. Methods: To confront these challenges, we established a sequential EV liquid-biopsy pipeline that integrates three complementary nanoengineered platforms for PDAC detection. First, a nanoplasmon-enhanced scattering (nPES) assay directly captures EVs from microliter plasma and quantifies tumor-enriched EphA2-positive EVs through dual antibody-nanoparticle coupling, simultaneously enabling the discovery of additional EV surface markers associated with PDAC. Second, we introduce FLARE (Fluctuation-enhanced simultaneous Labeling And Rapid Enrichment), which employs a low-frequency vibrating membrane and a peroxidase-mimicking nanozyme to synchronously enrich, label, and wash EVs in a single 45-minute operation, eliminating the need for multistep centrifugation and improving analytical performance. Third, we refined our plasmonic platform into a multiplex fluorescence assay that assembles plasmonic substrates in situ to enhance EV surface protein emission from unprocessed plasma, allowing simultaneous profiling of multiple PDAC-associated markers within one run. Results: The EphA2-EV nPES assay enabled highly sensitive quantification of tumor-derived EVs in both small-volume animal samples and patient plasma, distinguished PDAC from pancreatitis and healthy donors, and revealed dynamic EV changes before and after therapy. FLARE achieved more than ten-fold higher EV recovery than conventional methods while maintaining high labeling efficiency, and its amplified colorimetric output enabled smartphone-based quantification of multiple EV biomarkers, discriminating early-stage PDAC with an area under the curve of 0.95. The multiplex plasmon-enhanced fluorescence assay further improved signal-to-noise ratios for rare EV protein detection, enabling multi-marker readout from minimal plasma without prior EV isolation, facilitating personalized prognosis. Conclusions: By combining plasmonic scattering, vibration-assisted nanozyme enrichment, and plasmon-enhanced fluorescence, this EV-centered pipeline establishes a rapid, low-cost, and sample-sparing framework for PDAC liquid biopsy. Together, these platforms support scalable early detection, longitudinal treatment monitoring, and large-cohort translational studies across both clinical and preclinical settings. Citation Format: Tony Hu. Engineering next-generation EV diagnostics by integrating nanoplasmonic sensing and multiplex enrichment for early PDAC detection abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 2550.
Tony Hu (2026) studied this question.