Mutated Human P-Selectin Glycoprotein Ligand-1 and Viral Protein-1 of Enterovirus 71 Interactions on Au Nanoplasmonic Substrate for Specific Recognition by Surface-Enhanced Raman Spectroscopy

Round 1

Reviewer 1 Report

1. Yang et al showed the effect of the mutated protein and VP1 interactions by Raman spectroscopy. This method could be used for various mechanism detection.

2. I recommend this manuscript after providing proper references to mention background study and english editing.

Author Response

Dear Editor,

            We highly appreciate the reviewers’ comments that help to improve the quality of the revised manuscript. Their addressed questions have been carefully replied.

            Attached please find 4 files: the replies to reviewers, the list of changes, Supporting Data, and the revised manuscript.  

Thank you very much for your kind consideration to publish this paper in a regular issue.

Yours sincerely,

Jiunn-Der Liao

The corresponding author

Author Response File: Author Response.docx

Reviewer 2 Report

〇Comments

This manuscript by Kundan Sivashanmugan et al. addresses a validation of the interaction between mutated PSGL-1 and VP1. The authors described the method for validation of sulfated or mutated GST-PSGL-1 positions using SERS spectra, and the interaction of GST-PSGL-1s with VP1. It is important to show the position of tyrosine residues involved in the specific interaction. Additionally, this manuscript is clearly presented about the importance and the aim of work. However, the results are not well discussed, and the experiments are not enough. The results need to be improved with more explanations. I hope you to help with my comments.

1. Page7; About Fig.4, the authors described signal peaks at binding energies in text. However, it wasn’t clear whether we should pay attention to data in figure, so it is difficult for readers to understand it. The authors need to describe the explanation more clearly (e.g. it would be useful to include every commented data in Fig. 4.).
2. I wonder whether it would be better if each of the bottom of Figure 5 got an own number, (i) – (iv).
3. In Fig. 6, the authors described that mutated protein SERS intensity was reduced compared to that of the control due to the phenylalanine, and M2-5 showed enhanced SERS intensity. I fail to understand why this data is able to understand interactions with antibodies or VP1.
4. P10 Line345; The authors described that three mutated proteins indicate less interaction with VP1. However, it seems that a signal of V5 is almost same as that of V2 or V4 at 1600 cm^-1. How did the authors lead the conclusion? Additionally, it seems that the interaction toward virus much depend on 51 position, which inferred from Fig. 7c. Is there significance in other mutation, except for 51 position?
5. P2 L64; “used to” should be corrected without italic face.
6. P2 L80 and 84; “EV1” should be corrected as “EV71”.
7. P10 L329; Please check a unit of “1 um”.
8. The authors should describe official name for PBS at P4 L38 (not P10).
9. P10 L345; Please check “Figure 4(b)”. Is it not 4b but 7b?
10. I think “Marked SERS bands …” in legend of Figure 6 and 7 is explanation for (a), so I recommend “(b)” is transferred to place before “Relationship of relative …”. Moreover, the authors should describe figure legend for “Fig. 7c”.
11. There is no statement concerning Figure S4, Figure S5 and Table S5. Please check it.
12. Please check a text for Figure S4 in Supporting Information. Is it not “Figure S6(b)” but S4(b)?
13. It seems that there is no highlight for 1000 cm^-1 in Fig. S6. Please check it.

Author Response

Dear Editor,

            We highly appreciate the reviewers’ comments that help to improve the quality of the revised manuscript. Their addressed questions have been carefully replied.

            Attached please find 4 files: the replies to reviewers, the list of changes, Supporting Data, and the revised manuscript.  

Thank you very much for your kind consideration to publish this paper in a regular issue.

Yours sincerely,

Jiunn-Der Liao

The corresponding author

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

This manuscript by Kundan Sivashanmugan et al. addresses a validation of the interaction between mutated PSGL-1 and VP1. This second version of the manuscript has been improved. This article will be acceptable after some corrections have been done.

1. P2 L84; “EV1” is also corrected as “EV71”.
2. P10 L329; I think that the correct unit is “1 μM” not “1 μm”…
3. ‘Figure S4', 'Figure S5' and 'Table S5’ are missing in the text. Please check it.