Peer Review History

Original SubmissionJune 4, 2025
Decision Letter - Rui Tada, Editor

Dear Dr. Sato,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by Sep 05 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

  • A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
  • A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
  • An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Rui Tada, Ph.D.

Academic Editor

PLOS ONE

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. We note that there is identifying data in the Supporting Information file <20250522_Supplement Information _Haruya_Sato.docx>. Due to the inclusion of these potentially identifying data, we have removed this file from your file inventory. Prior to sharing human research participant data, authors should consult with an ethics committee to ensure data are shared in accordance with participant consent and all applicable local laws.

Data sharing should never compromise participant privacy. It is therefore not appropriate to publicly share personally identifiable data on human research participants. The following are examples of data that should not be shared:

-Name, initials, physical address

-Ages more specific than whole numbers

-Internet protocol (IP) address

-Specific dates (birth dates, death dates, examination dates, etc.)

-Contact information such as phone number or email address

-Location data

-ID numbers that seem specific (long numbers, include initials, titled “Hospital ID”) rather than random (small numbers in numerical order)

Data that are not directly identifying may also be inappropriate to share, as in combination they can become identifying. For example, data collected from a small group of participants, vulnerable populations, or private groups should not be shared if they involve indirect identifiers (such as sex, ethnicity, location, etc.) that may risk the identification of study participants.

Additional guidance on preparing raw data for publication can be found in our Data Policy (https://journals.plos.org/plosone/s/data-availability#loc-human-research-participant-data-and-other-sensitive-data) and in the following article: http://www.bmj.com/content/340/bmj.c181.long.

Please remove or anonymize all personal information (Name), ensure that the data shared are in accordance with participant consent, and re-upload a fully anonymized data set. Please note that spreadsheet columns with personal information must be removed and not hidden as all hidden columns will appear in the published file.

3. PLOS requires an ORCID iD for the corresponding author in Editorial Manager on papers submitted after December 6th, 2016. Please ensure that you have an ORCID iD and that it is validated in Editorial Manager. To do this, go to ‘Update my Information’ (in the upper left-hand corner of the main menu), and click on the Fetch/Validate link next to the ORCID field. This will take you to the ORCID site and allow you to create a new iD or authenticate a pre-existing iD in Editorial Manager.

4. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. 

5. If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. 

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Partly

**********

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

Reviewer #1: Yes

Reviewer #2: N/A

Reviewer #3: N/A

**********

3. Have the authors made all data underlying the findings in their manuscript fully available??>

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: No

Reviewer #3: No

**********

4. Is the manuscript presented in an intelligible fashion and written in standard English??>

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

**********

Reviewer #1: This study reports on the development of a novel methodology for conjugation of reporter molecules to antibody fragments under mild conditions, which is more straightforward compared to the previously developed approaches.

The study is well performed and the manuscript is well written. I only have minor questions:

In Figure 1, n= should be specified to what was used in this study.

Table 1, was the measurement only performed once? What is n= and SD=?

SI page 5 “LC-MS/MS analysis of tryptic digest of Fab. For LC-MS/MS analysis, 2 μL of 1 M Tris-HCl solution (pH 8.5) was added to 20 μL of 0.25 mg/mL Fab solution in milli-Q H2O containing 8 M urea (grade, maker).” - 8 M urea (grade, maker) should be added.

Reviewer #2: This paper describes the method for the preparation of Ab fragment-reporter conjugate by the two-step reaction: first step is the introduction of bifunctional linker to the Q tag attached to Ab via side chain amide bond. Subsequently, the thiol group of the linker was reacted with maleimide linked reporter. The advantage of this method is that the thiol group can be made free during the reaction with Ab, which realizes the direct coupling with the reporter without the deprotection step. Conceptually, the paper represents no surprising idea, as the use of Q-tag and transglutaminase has already reported and it is natural to think that the protection of thiol can be omitted in this reaction. However, the author successfully proved that the reaction indeed proceeded as suggested using several model proteins and this reviewer thinks that the paper may be suitable for publication with the following modifications.

1. SH-PEG-NH2 should be HS-PEG-NH2. In addition, in Fig. 1 PEG linker should be NH2....SH.

2. In page 3, line 8, making it difficult to control the stoichiometry-> The compound prepared in this paper is also heterogeneous, although at reporter part which does not affect the binding affinity. Still, it would be difficult to precisely control the stoichiometry to obtain the target conjugates. It is better to revise the expression.

3. Page 5, line 6: speculating that the steric hindrance... -> I do not understand the meaning. Please revise.

4. Page 8, line 5, page 13 line 3 from the bottom: likely owing to conjugate loss -> Please add the brief result of cation exchange data.

5. There is no experimental procedure for Table 1. Please add the errors of the KD value.

6. There are not MS data for the final conjugates. Please add them.

7. Do the authors intend to use this conjugate as a trimer? If so, it is better to optimize the yield of the trimer, as the purification by SEC is always necessary, which is not so efficient.

Reviewer #3: The authors have described a two-step chemoenzymatic method to generate antibody conjugate involving transglutaminase followed by a thio-maleimide Michael addition reaction. The key of this study is the demonstration of a long-chain thiol-PEG-amine (SH-PEG) linker is relatively stable than the short-chain thiol-PEG-amine during the transglutaminase mediated reactions, thus, to avoid the traditional thio group protection-deprotections procedure. Overall, the study provides a useful method for facilitating thio-maleimide Michael addition approach in antibody conjugate. However, several improvements should be made for before publication.

1. In Figure 2, the monomer is detected exclusively as a singly‑charged ion, while the dimer appears only as a doubly‑charged species. Although the dimer signal is minor (~2%, as stated by the authors), it would be more informative to include the mono‑charged form of the dimer as well, if it is present. This would help validate the assignment and improve the completeness of the mass spectrometric analysis.

2. Page 13, line 7, Fabp24 should not be purified by Protein G chromatography, unless it is engineered to be part of a fusion protein that binds to Protein G, which is not typical. It maybe a reason why the yield of Alexa488-PEG2k-Fabp24 is low (33%). Instead, Protein L should be applied.

3. Page 15, line 10: SPR spectra should be provided.

**********

what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy

Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step.

https://doi.org/10.1371/journal.pone.0333359.r002
Revision 1
Author Response

Rebuttal letter to reviewer’s Comments

Thank you very much for reviewing our manuscript. We appreciate your useful comments and advice. We describe the replies to your comments and advice as blue letters.

In addition to the changes made in response to your comments, we have corrected the author's postal code (Manuscript, p1, line 3 from the bottom), which was incorrect. Furthermore, we have revised an erroneous description in Figure S7 (Supplementary Materials, p20, lines 1-2 and line 7). We appreciate your understanding in these corrections.

Reviewer #1: This study reports on the development of a novel methodology for conjugation of reporter molecules to antibody fragments under mild conditions, which is more straightforward compared to the previously developed approaches.

The study is well performed, and the manuscript is well written. I only have minor questions:

In Figure 1, n= should be specified to what was used in this study.

Thank you for pointing this out. Figure 1 is conceptual; thus we emphasized the number n (from 39 to 55) in another place in the text (p6, line 9).

Table 1, was the measurement only performed once? What is n= and SD

Thank you for your comments. The KD values for each sample were evaluated based on one or two assays (n= 1 or 2) using five different concentrations of analyte. To clarify this point, we have included the SPR experimental results and sensorgram figures (p7, lines; p15, Figure S5) in the Supplementary Information. We have also detailed the description of these results to include the equivalence of antigen-binding activity between unmodified and modified Fabp24, as follows (p7 from line 8 from the bottom to p8 line 2, in bold).

SPR analyses of Fabp24, HS-PEG2k-Fabp24, and Alexa488-PEG-Fabp24 against HIV-1 p24 protein.

Fab and its derivatives were immobilized on a sensor chip and injected at five different concentrations of recombinant HIV-1 p24 protein (Figure S5). As expected, HIV-1 p24 protein bound specifically to the Fabs. In contrast, no affinity for HIV-1 p24 protein was observed on sensor chips without the Fab (data not shown). The sensorgram profiles of HS-PEG2k-Fabp24 and Fabp24 (Figure S5A and S5B) and Alexa488-PEG2k-Fabp24 and Fabp24 (Figure S5C and S5D), each evaluated in a single batch of experiments, were highly similar, and their KDs were equivalent. Based on these results, the antigen-binding activity of modified Fabp24 and Fabp24 were considered equivalent.

Additionally, we added explanations to the footnote in Table 1 (p10) and the Materials and Methods section in the text (p20, line 6).

p10, Table 1 footnote: *Average value of 2 independent data (For Fabp24)

p20, line 5 (Bold): The interaction was analysed using a 1:1 binding model with Biacore™ T200 evaluation software in one or two assays using five different analyte concentrations.

SI page 5 “LC-MS/MS analysis of tryptic digest of Fab. For LC-MS/MS analysis, 2 μL of 1 M Tris-HCl solution (pH 8.5) was added to 20 μL of 0.25 mg/mL Fab solution in milli-Q H2O containing 8 M urea (grade, maker).” - 8 M urea (grade, maker) should be added.

We have added the information in the Supplement (p5, line 3).

Reviewer #2: This paper describes the method for the preparation of Ab fragment-reporter conjugate by the two-step reaction: first step is the introduction of bifunctional linker to the Q tag attached to Ab via side chain amide bond. Subsequently, the thiol group of the linker was reacted with maleimide linked reporter. The advantage of this method is that the thiol group can be made free during the reaction with Ab, which realizes the direct coupling with the reporter without the deprotection step. Conceptually, the paper represents no surprising idea, as the use of Q-tag and transglutaminase has already reported and it is natural to think that the protection of thiol can be omitted in this reaction. However, the author successfully proved that the reaction indeed proceeded as suggested using several model proteins and this reviewer thinks that the paper may be suitable for publication with the following modifications.

1. SH-PEG-NH2 should be HS-PEG-NH2. In addition, in Fig. 1 PEG linker should be NH2....SH.

Thank you for your advice. We have altered the nomenclature of SH-PEG and to HS-PEG in our revised manuscript. Further details are provided in the revised manuscript. In Figure 1, we also revised the description of linker’s structure as NH2-PEG-SH.

2. In page 3, line 8, making it difficult to control the stoichiometry-> The compound prepared in this paper is also heterogeneous, although at reporter part which does not affect the binding affinity. Still, it would be difficult to precisely control the stoichiometry to obtain the target conjugates. It is better to revise the expression.

We appreciate your advice. The expression has been revised in the text �p3, line 8-9�, as indicated in bold below.

In these reactions, targeting specific residues among the multiple lysine and cysteine residues of the antibody is challenging, making it difficult to limit the stoichiometry to a certain range, leading to the synthesis of heterogeneous conjugates with low antigen-binding activity.

3. Page 5, line 6: speculating that the steric hindrance... -> I do not understand the meaning. Please revise.

Thank you for your comments. We have revised the description in the text (p5, line 6-9), as indicated in bold below.

This strategy was achieved using an HS-PEG linker based on the speculation that the unique structure of this molecule creates spatial constraints, which prevent collisions between the thiol groups at the end of each molecule to inhibit the formation of S-S bonds.

4. Page 8, line 5, page 13 line 3 from the bottom: likely owing to conjugate loss -> Please add the brief result of cation exchange data.

Thank you for your indication. We have added the description about conjugate loss and brief results of cation exchange data in the revised text, as described below.

p8, line 6 (Bold):

likely owing to conjugate loss during affinity chromatography and concentration steps.

P14, line 4 from the bottom (Bold)

with Protein G and concentration step by using ultrafiltration to remove the unreacted HS-PEG linker.

p14, line 2 from the bottom (Bold):

Instead of this chromatography, we have obtained high yields by using cation exchange chromatography (over 80%, data not shown) for the upcoming scaled-up conjugation process.

5. There is no experimental procedure for Table 1. Please add the errors of the KD value.

Thank you for your comments. The KD values for each sample were evaluated based on one or two assays (n= 1 or 2) using five different concentrations of analyte. To clarify this point, we have included the SPR experimental results and sensorgram figures (p7, lines; p15, Figure S5) in the Supplementary Information. We have also detailed the description of these results to include the equivalence of antigen-binding activity between unmodified and modified Fabp24, as follows (p7 from line 8 from the bottom to p8 line 2, in bold).

SPR analyses of Fabp24, HS-PEG2k-Fabp24, and Alexa488-PEG-Fabp24 against HIV-1 p24 protein.

Fab and its derivatives were immobilized on a sensor chip and injected at five different concentrations of recombinant HIV-1 p24 protein (Figure S5). As expected, HIV-1 p24 protein bound specifically to the Fabs. In contrast, no affinity for HIV-1 p24 protein was observed on sensor chips without the Fab (data not shown). The sensorgram profiles of HS-PEG2k-Fabp24 and Fabp24 (Figure S5A and S5B) and Alexa488-PEG2k-Fabp24 and Fabp24 (Figure S5C and S5D), each evaluated in a single batch of experiments, were highly similar, and their KDs were equivalent. Based on these results, the antigen-binding activity of modified Fabp24 and Fabp24 were considered equivalent.

Additionally, we added explanations to the footnote in Table 1 (p10) and the Materials and Methods section in the text (p20, line 6).

p10, Table 1 footnote: *Average value of 2 independent data (For Fabp24)

p20, line 6 (Bold): The interaction was analysed using a 1:1 binding model with Biacore™ T200 evaluation software in one or two assays using five different analyte concentrations.

6. There are not MS data for the final conjugates. Please add them.

Thank you for your comment. As you pointed out, measuring the mass of the conjugate is very important. Unfortunately, these molecules are too large (>310 kDa) for our mass spectrometer, and we were unable to measure their masses.

7. Do the authors intend to use this conjugate as a trimer? If so, it is better to optimize the yield of the trimer, as the purification by SEC is always necessary, which is not so efficient.

We do not have a plan to use the (Fab-PEG)3-PE conjugate as a reagent for in vitro research or IVD reagents because its yield is too low. However, since the composition of HS-PEG-Fab conjugates with PE is highly reproducible under constant reaction conditions, we plan to use the total product fraction for our research and IVDs.

Reviewer #3: The authors have described a two-step chemoenzymatic method to generate antibody conjugate involving transglutaminase followed by a thio-maleimide Michael addition reaction. The key of this study is the demonstration of a long-chain thiol-PEG-amine (SH-PEG) linker is relatively stable than the short-chain thiol-PEG-amine during the transglutaminase mediated reactions, thus, to avoid the traditional thio group protection-deprotections procedure. Overall, the study provides a useful method for facilitating thio-maleimide Michael addition approach in antibody conjugate. However, several improvements should be made for before publication.

1. In Figure 2, the monomer is detected exclusively as a singly charged ion, while the dimer appears only as a doubly charged species. Although the dimer signal is minor (~2%, as stated by the authors), it would be more informative to include the mono charged form of the dimer as well, if it is present. This would help validate the assignment and improve the completeness of the mass spectrometric analysis.

Unfortunately, we were unable to detect the singly charged ion of the dimerized PEG molecule by MS analysis due to its high m/z. Thus, we estimated the amount of the molecule by the doubly charged ion.

2. Page 13, line 7, Fabp24 should not be purified by Protein G chromatography, unless it is engineered to be part of a fusion protein that binds to Protein G, which is not typical. It may be a reason why the yield of Alexa488-PEG2k-Fabp24 is low (33%). Instead, Protein L should be applied.

Thank you for your comment. We have made a preliminary comparison of the yield of HS-PEG-modified Fab using Protein G and Protein L chromatography, and the recoveries were similar. Since Protein G resin is cheaper than Protein L resin, we selected the Protein G column to purify the PEG conjugates and remove the unmodified HS-PEG linkers.

3. Page 15, line 10: SPR spectra should be provided.

We have included the SPR experimental results and sensorgram figures (p7, lines; p15, Figure S5) in the Supplementary Information. We have also detailed the description of these results to include the equivalence of antigen-binding activity between unmodified and modified Fabp24, as follows (p7 line 8-1 from the bottom to p8 line 2, in bold).

SPR analyses of Fabp24, HS-PEG2k-Fabp24, and Alexa488-PEG-Fabp24 against HIV-1 p24 protein.

Fab and its derivatives were immobilized on a sensor chip and injected at five different concentrations of recombinant HIV-1 p24 protein (Figure S5). As expected, HIV-1 p24 protein bound specifically to the Fabs. In contrast, no affinity for HIV-1 p24 protein was observed on sensor chips without the Fab (data not shown). The sensorgram profiles of HS-PEG2k-Fabp24 and Fabp24 (Figure S5A and S5B) and Alexa488-PEG2k-Fabp24 and Fabp24 (Figure S5C and S5D), each evaluated in a single batch of experiments, were highly similar, and their KDs were equivalent. Based on these results, the antigen-binding activity of modified Fabp24 and Fabp24 were considered equivalent.

Attachments
Attachment
Submitted filename: 20250901_Response to reviewers.docx
https://doi.org/10.1371/journal.pone.0333359.r003
Decision Letter - Rui Tada, Editor

Efficient Two-Step Chemoenzymatic Conjugation of Antibody Fragments with Reporter Compounds by a Specific thiol-PEG-amine Linker, HS-PEG-NH2

PONE-D-25-30206R1

Dear Dr. Sato,

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.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager®  and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact billing support .

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Rui Tada, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewer #1:

Reviewer #2:

Reviewer #3:

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions??>

Reviewer #1: (No Response)

Reviewer #2: (No Response)

Reviewer #3: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously? -->?>

Reviewer #1: (No Response)

Reviewer #2: (No Response)

Reviewer #3: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available??>

The PLOS Data policy

Reviewer #1: (No Response)

Reviewer #2: (No Response)

Reviewer #3: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English??>

Reviewer #1: (No Response)

Reviewer #2: (No Response)

Reviewer #3: Yes

**********

Reviewer #1: (No Response)

Reviewer #2: (No Response)

Reviewer #3: The authors have properly responded to my comments. I recommend the acceptance of the current manuscript.

**********

what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy

Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

**********

https://doi.org/10.1371/journal.pone.0333359.r004
Formally Accepted
Acceptance Letter - Rui Tada, Editor

PONE-D-25-30206R1

PLOS ONE

Dear Dr. Sato,

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team.

At this stage, our production department will prepare your paper for publication. This includes ensuring the following:

* All references, tables, and figures are properly cited

* All relevant supporting information is included in the manuscript submission,

* There are no issues that prevent the paper from being properly typeset

You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps.

Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing.

If we can help with anything else, please email us at customercare@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Rui Tada

Academic Editor

PLOS ONE

https://doi.org/10.1371/journal.pone.0333359.r005

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .