2026-10-11T13:36:52Z https://api.figshare.com/v2/oai

oai:figshare.com:article/33994230 2026-09-25T09:06:56Z category_1 category_4 category_146 category_7 category_14 category_21 category_873 category_64 category_135 portal_63 item_type_2 month_year_09_2026

Molecular Programming of Steric Tags for Controlled Nanopore Translocation toward Precision Clinical Diagnostic Assistance Yiheng Liu (11475805) Shijun Lin (2087206) Minglun Li (28228) Hao Fang (69476) Zhicheng Zhang (490080) Ruwei Wei (13176404) Yan Zhang (8098) Jun Dai (435229) Tao Liu (10785) Fan Xia (219575) Xiaoding Lou (1406035) Biophysics Biochemistry Space Science Medicine Molecular Biology Biotechnology Chemical Sciences not elsewhere classified Cancer Computational Biology stochastic thermal motion precisely defined dimensions magnitude dynamic range klk3 activity analysis general chemical framework conventional detection bandwidths confined nanoscale environments clinical serum samples additional protease target peptide residence time modular probe design programming molecular behavior functional nanopore sensing 2 nm constriction enabled slow translocation molecular programming rational design peptide substrates enabled classification work establishes urothelial carcinoma trans ) tag size systematic variation steric tags precise resolution physical limitation often limited nanopore chemistry molecule events matrix metalloproteinases m2mspa nanopore learning classifier fundamental challenge flexible peptides derived measurements concept extension actively reshape 9 ). The precise resolution of single-molecule events is often limited by the stochastic thermal motion of analytes, which traverse nanoscale sensing zones too rapidly for conventional detection bandwidths. This remains a fundamental challenge in nanopore chemistry, particularly for small and flexible peptides. Here, we report a chemically intuitive molecular programming strategysteric blockage-enabled slow translocation (SLOW-Trans)that overcomes this physical limitation through the rational design of sterically demanding probe molecules. By site-specifically conjugating rigid aromatic steric tags with precisely defined dimensions to peptide substrates, we actively reshape the translocation energy landscape within the ∼1.2 nm constriction of the M2MspA nanopore. Systematic variation of tag size and rigidity enables deterministic control over peptide residence time, transforming transient stochastic events into stable and highly discriminable current signatures. Using this strategy, we achieve a six-order-of-magnitude dynamic range (0.001–1000 ng/mL) for matrix metalloproteinases (MMP-1, -2, and -9). In a cohort of 231 clinical urine samples, the nanopore-derived MMP activity profiles showed excellent agreement with ELISA-derived measurements and, when integrated with a machine-learning classifier, enabled classification of urothelial carcinoma with 96.2% accuracy. As a proof-of-concept extension, the modular probe design was further adapted for PSA/KLK3 activity analysis in clinical serum samples, supporting the feasibility of extending the approach to an additional protease target. This work establishes a general chemical framework for programming molecular behavior in confined nanoscale environments, bridging molecular design principles with functional nanopore sensing. 2026-09-25T00:00:00Z Dataset Media 10.1021/jacs.6c15448.s002 https://figshare.com/articles/media/Molecular_Programming_of_Steric_Tags_for_Controlled_Nanopore_Translocation_toward_Precision_Clinical_Diagnostic_Assistance/33994230 CC BY-NC 4.0