Immobilized Fluorescent Probes for Simultaneous Multiple Protease Detection
Round 1
Reviewer 1 Report
Please find a file report enclosed
Comments for author File: 
Comments.pdf
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Reviewer 2 Report
The manuscript by Milićević and Hlaváč describes a new assay for protease detection. Overall, it is a high quality manuscript that provides sufficient experimental details and thorough discussion. The manuscript is well written.
The recommendation is to accept for publication in a current form.
Author Response
Thank you for the reviewing our manuscript.
Reviewer 3 Report
In this manuscript Milićević et al have demonstrated application of simultaneous detection of two cleavable enzymes on solid support. The enzymes were connected to two enzymes by PEG linkers. Using a single excitation source the authors were able to determine the cleavage efficiency of different enzymes. As such, the reviewer did not come across any major issues in the manuscript and the work. The work is well presented and would be of interest to the audience of chemo sensors.
Minor Changes.
- Figure 3A and B. Why is does not go to baseline after lambda max? Fig3A, the absorbance spectra should be repeated because it has reached the saturation point.
- Will the peptide cleavage be effective at different basic pH? It would be worthwhile for the reviewers to run the assay in different pHs.
- What was the solvent and the temperature at which the authors carried out the study?
- The reason for superiority of Rink Amide resin over Tentagel XV and Chemmatrix resin should be discussed in the article.
- Figure 4 and 5. It seems that the dye gets released non-specifically over time in control experiments. The reasons for this should be discussed in the manuscript.
- It is advised that the authors run experiments on stability of the dyes and dye conjugates (without resin) to determine the non-specific cleavage over time.
Author Response
Please, see the attachment
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Dear Editor,
after a careful evaluation of the new version of the manuscript it is reviewer's opinion that the authors have properly replied to all comments providing the paper with the required changes. Now the manuscript has improved and, for this reason, its publication is suggested.
Best regards
