PONE-D-24-05407Comparative analyses of adsorbed circulating proteins in the PMMA and PES hemodiafilters in patients on predilution online hemodiafiltrationPLOS ONE

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Additional Editor Comments:

This manuscript is well written, and should meet the publication criteria with a few minor edits in response to the two reviewers

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

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Reviewer #1: Partly

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: No

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

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Reviewer #1: The authors clarified, in patients undergoing online HDF (Hemodiafiltration), what substances are adsorbed by PES (Polyethersulfone) and PMMA (Polymethylmethacrylate) membranes respectively, using mass spectrometry. The method of identifying substances adsorbed on the dialysis membrane is considered to be without question and valid. However, in dialysis, the removal of specific substances is significantly influenced not only by adsorption to the dialysis membrane but also by excretion into the filtrate. In this study, the discussion focuses solely on adsorption, and it is deemed meaningless to argue the clinical significance based on the difference in the amount of substances adsorbed.

Reviewer #2: This manuscript by Shoriful Islam et al. provides a succinct study of proteins absorbed by dialysis membranes in the treatment of patients with kidney disease. The authors assess three patients using two different membrane materials, and analyse the absorbed proteins using standard SDS-PAGE and LC-MS/MS proteomics. They were able to find differences in protein absorption between the two membrane materials.

The experimental procedure has been well written with all the relevant details for the proteomics analysis that will allow reproducibility. MS/MS traces have been included in the supporting information, and details of the Mascot searches are also included.

I do have some questions however, before I can recommend publication.

- What was the rationale for first placing the patients with the PES membrane and then switching to the PMMA membrane, would that not skew the results for the PMMA membrane as the PES might absorb proteins first?

- There are some instances where a PS membrane is discussed, is this a typo for PES, or a different type of membrane?

- Figure 2 quantifies the levels of proteins absorbed onto the different membranes. How was this quantification performed? I couldn't find these details in the experimental or in the results section. Also, was any quantitative analysis of band intensity in the gels performed, or was any discussion on band intensity purely qualitative? I would suggest densitometry analysis through ImageJ for example would be more accurate here.

- The second sentence of the abstract is vague for those not in the field - is the hollow fiber in reference to the dialysis membrane?

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Reviewer #1: Yes: Noriaki Iino

Reviewer #2: No

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Comparative analyses of adsorbed circulating proteins in the PMMA and PES hemodiafilters in patients on predilution online hemodiafiltration

PONE-D-24-05407R1

Dear Dr. Setou,

Thank you for your detailed response to the reviewer comments. We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Robert Chapman, Ph.D.

Academic Editor

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