The manuscript presents a promising MoS₂/silk fibroin FET biosensor for cTnI detection, but several important issues remain regarding novelty, LOD validation, sensing mechanism, reproducibility, and clinical applicability. The manuscript may become suitable for publication after the authors adequately address these major concerns.

1. • Could the authors clearly clarify the novelty of the present MoS₂/silk fibroin biosensor compared with their previous reports on biopolymer-encapsulated 2D FET biosensors, particularly the previously reported Cs₂AgBiBr₆-based cTnI sensor? The specific scientific advance of the present work should be explicitly established.
2. • How was the reported LOD of 1.14 pg/mL calculated? Please provide the calibration equation, slope, standard deviation of the blank, number of independent measurements, and the statistical criterion used for LOD determination.
3. • How can the authors experimentally support an LOD of 1.14 pg/mL when the calibration data shown in Fig. 5 appear to start at 10 pg/mL? Please provide experimental measurements below 10 pg/mL rather than relying on extrapolation from higher concentrations.
4. • Could the authors provide appropriate control experiments to distinguish the individual contributions of MoS₂, yolk-shell AuNR@Au/Ag nanostructures, and antibody functionalization to the overall sensing response?
5. • What is the underlying mechanism responsible for the change in the IDS–VDS slope upon cTnI binding? The authors should provide stronger experimental evidence to distinguish charge transfer, electrostatic gating, carrier-density modulation, and/or contact-resistance effects.
6. • Could the authors demonstrate the reproducibility of the sensor fabrication and response using multiple independently fabricated MoS₂ FET devices and different synthesis batches? Please clarify whether the reported n = 3 represents independent devices or repeated measurements of the same device.
7. • Does the approximately 98 nm silk fibroin layer itself influence the electrical properties and sensing response of the MoS₂ FET? Appropriate controls should be provided to separate the protective effect of silk from its possible influence on electrostatic screening and charge transport.
8. • What direct evidence demonstrates that silk fibroin preserves the biological activity of the immobilized cTnI antibody, rather than simply protecting the underlying sensor components from thermal degradation?
9. • Could the authors provide sensing performance in real human serum or plasma samples? Since the manuscript emphasizes clinical AMI diagnostics and claims applicability to human plasma, recovery, matrix-effect, and ideally comparison with a standard clinical assay are necessary to substantiate these claims.
10. • Are the concentrations of CRP, myoglobin, hemoglobin, glucose, and uric acid used in the selectivity experiments clinically relevant? Please justify the selected concentrations and consider testing additional cardiovascular biomarkers under realistic concentration ratios.
11. • Could the authors provide a comprehensive comparison with state-of-the-art cTnI biosensors, including LOD, linear range, response time, sample matrix, selectivity, thermal/storage stability, and device architecture? This comparison is necessary to demonstrate the practical and scientific advantage of the proposed platform.
12. • Could the authors reconsider the broad claims regarding “point-of-care” and “practical applications,” given that the current platform requires CVD growth, lithographic fabrication, synthesis of complex yolk-shell nanostructures, antibody conjugation, and multiple surface-functionalization steps? The practical advantages and remaining technological limitations should be discussed more critically.
13. In the section of Introduction, some related works for CTnI detection should be discussed or cited, ACS Applied Nano Materials 2023, 6 (20), 19239-19248, Sensors and Actuators B: Chemical 2023, 397, 134668. Electrochimica Acta 559 (2026) 148538 (10.1016/j.electacta.2026.148538)
14. The analytical validation would benefit from stronger statistical support. For example, recovery results could be evaluated using a paired t-test to determine whether the measured concentrations significantly differ from the spiked values. In addition, the reported %RSD values should be interpreted against established precision criteria, such as those provided by AOAC or ICH guidelines, or by comparison with values reported in similar electrochemical biosensor studies. Including these statistical justifications and relevant references would strengthen the reliability of the validation results. Journal of The Electrochemical Society 2025, 172, 077511 (10.1149/1945-7111/adf2e5), Microchimica Acta 192 (2), 113, 2025 (10.1007/s00604-025-06968-7), Electrochimica Acta 559 (2026) 148538 (10.1016/j.electacta.2026.148538), Journal of Electroanalytical Chemistry 992 (2025) 119215 (10.1016/j.jelechem.2025.119215), Electrochimica Acta, 149396, 574, 2026 (10.1016/j.electacta.2026.149396)
15. It is recommended that the authors evaluate both intraday and interday precision of the proposed method and provide the corresponding statistical analysis (e.g., Student’s t-test at a 95% confidence level, α = 0.05). Including such results will help to demonstrate the robustness and reliability of the developed biosensor. This point should be supported with an appropriate reference. Microchemical Journal, 205, 2024, 111382, (10.1016/j.microc.2024.111382), Microchimica Acta 2024, 191 (9), 546 (10.1007/s00604-024-06622-8), Microchemical Journal 220, 2026, 116602 (10.1016/j.microc.2025.116602),