A label-free optical biosensor based on porous silicon (PSi) was developed for the high-sensitivity detection of human cardiac troponin T (cTnT), a key biomarker for the early diagnosis of myocardial infarction (MI). To reduce synthesis costs while maintaining binding efficiency, a truncated peptide nucleic acid (PNA) probe was designed in silico starting from a 40-base-long wild-type sequence. Computational screening identified a 12-base candidate, whose binding affinity was experimentally validated by Western blot analysis. The freshly etched PSi surface was passivated and functionalized via mild thermal hydrosilylation with 10-undecenoic acid to introduce carboxylic groups, enabling covalent PNA immobilization through the carbodiimide chemistry. Surface functionalization and stability in aqueous media were validated using both label-free optical reflectance and fluorescence spectroscopy. Probe conjugation was optimized at pH 5.5, resulting in a surface density of 1.12 ± 0.30 pmol cm−2. The biosensor exhibited reproducible, concentration-dependent responses to cTnT in the 0.02–0.16 ng mL−1 range, with a limit of detection of 0.030 ± 2.0 × 1 0 −5 ng mL−1. This performance is clinically relevant, as cTnT levels in healthy individuals are typically <0.01 ng mL−1, while MI patients often exceed 10 ng mL−1, reaching >100 ng mL−1 in severe cases.
Truncated PNA-functionalized porous silicon biosensor for low-cost and early detection of troponin T in myocardial infarction
Nocerino, Valeria;
2026-01-01
Abstract
A label-free optical biosensor based on porous silicon (PSi) was developed for the high-sensitivity detection of human cardiac troponin T (cTnT), a key biomarker for the early diagnosis of myocardial infarction (MI). To reduce synthesis costs while maintaining binding efficiency, a truncated peptide nucleic acid (PNA) probe was designed in silico starting from a 40-base-long wild-type sequence. Computational screening identified a 12-base candidate, whose binding affinity was experimentally validated by Western blot analysis. The freshly etched PSi surface was passivated and functionalized via mild thermal hydrosilylation with 10-undecenoic acid to introduce carboxylic groups, enabling covalent PNA immobilization through the carbodiimide chemistry. Surface functionalization and stability in aqueous media were validated using both label-free optical reflectance and fluorescence spectroscopy. Probe conjugation was optimized at pH 5.5, resulting in a surface density of 1.12 ± 0.30 pmol cm−2. The biosensor exhibited reproducible, concentration-dependent responses to cTnT in the 0.02–0.16 ng mL−1 range, with a limit of detection of 0.030 ± 2.0 × 1 0 −5 ng mL−1. This performance is clinically relevant, as cTnT levels in healthy individuals are typically <0.01 ng mL−1, while MI patients often exceed 10 ng mL−1, reaching >100 ng mL−1 in severe cases.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.


