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Early detection of pancreatic ductal adenocarcinoma by single-molecule profiling of pancreatic enzyme activities

Sakamoto, S.; Hiraide, H.; Hatakeyama, J.; Kobayashi, T.; Takahashi, K.; Inagaki, T.; Takagi-Niidome, S.; Minoda, M.; Kawaguchi, T.; Shibayama, T.; Iwakura, N.; Kawana, M.; Mizuno, T.; Kawaguchi, M.; Nakagawa, H.; Fujita, K.; Urano, Y.; Masuda, A.; Ikemoto, J.; Ishii, Y.; Oka, S.; Hanada, K.; Kodama, Y.; Komatsu, T.; Kagami, Y.

2026-06-09 biochemistry
10.64898/2026.06.05.730533 bioRxiv
Show abstract

Pancreatic ductal adenocarcinoma (PDAC) remains a leading cause of cancer-related mortality, largely due to diagnosis at advanced stages. Early detection through minimally invasive liquid biopsy holds promise for improving patient outcomes. Here, we report a blood-based liquid biopsy platform based on single-molecule enzyme activity profiling (SEAP), which detects proteoform-level alterations in circulating pancreatic enzymes at single-molecule resolution. Using a tissue-centric biomarker discovery strategy, we identified activity signatures of pancreas-specific digestive enzymes associated with PDAC. A combinatorial classifier detected PDAC (stage I-IV) with 95.5% specificity and 75.0% sensitivity (74.2% for stage I-II) across 690 blood samples collected from multiple hospitals and biobanks. Performance was further validated in an independent cohort enriched for early-stage disease, where 54.2% (13/24) of stage IA, 64.3% (9/14) of stage IB, and 21.4% (3/14) of stage 0 lesions were classified as positive. These findings support the clinical potential of SEAP for early detection of PDAC.

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