Thermostable chaperone-based polypeptide biosynthesis: Enfuvirtide model product quality and protocol-related impurities

Vladimir Zenin; Andrey Tsedilin; Maria Yurkova; Andrey Siniavin; Alexey Fedorov

doi:10.1371/journal.pone.0286752

Peer Review History

Original SubmissionNovember 3, 2022
6 Feb 2023 Decision Letter - Richard Ali, Editor PONE-D-22-30362Thermostable chaperone-based polypeptide biosynthesis: enfuvirtide model product quality and protocol-related impuritiesPLOS ONE Dear Dr. Zenin, 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. The manuscript has been evaluated by two reviewers, and their comments are available below. The reviewers have raised a number of concerns that need attention. In particular, due to technical considerations, they have suggested that some experiments require repeating (protein quantification). Furthermore, the reviewers have suggested additional data can strengthen the manuscript. And finally, the reviewers have concerns around grammatical errors, and thus suggest copyediting for English language and grammar. Could you please revise the manuscript to carefully address the concerns raised? Please submit your revised manuscript by Mar 20 2023 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. Please include the following items when submitting your revised manuscript: 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, Richard Ali Staff 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 noticed you have some minor occurrence of overlapping text with the following previous publication(s), which needs to be addressed: - \\\\https://journals.lww.com/aidsonline/Fulltext/2019/01020/Structural_and_functional_characterization_of.1.aspx? In your revision ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the methods section. Further consideration is dependent on these concerns being addressed. 3. Thank you for stating the following financial disclosure: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” At this time, please address the following queries: a) Please clarify the sources of funding (financial or material support) for your study. List the grants or organizations that supported your study, including funding received from your institution. b) State what role the funders took in the study. If the funders had no role in your study, please state: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” c) If any authors received a salary from any of your funders, please state which authors and which funders. d) If you did not receive any funding for this study, please state: “The authors received no specific funding for this work.” Please include your amended statements within your cover letter; we will change the online submission form on your behalf. 4. We note that you have indicated that data from this study are available upon request. PLOS only allows data to be available upon request if there are legal or ethical restrictions on sharing data publicly. For more information on unacceptable data access restrictions, please see http://journals.plos.org/plosone/s/data-availability#loc-unacceptable-data-access-restrictions. In your revised cover letter, please address the following prompts: a) If there are ethical or legal restrictions on sharing a de-identified data set, please explain them in detail (e.g., data contain potentially sensitive information, data are owned by a third-party organization, etc.) and who has imposed them (e.g., an ethics committee). Please also provide contact information for a data access committee, ethics committee, or other institutional body to which data requests may be sent. b) If there are no restrictions, please upload the minimal anonymized data set necessary to replicate your study findings as either Supporting Information files or to a stable, public repository and provide us with the relevant URLs, DOIs, or accession numbers. For a list of acceptable repositories, please see http://journals.plos.org/plosone/s/data-availability#loc-recommended-repositories. We will update your Data Availability statement on your behalf to reflect the information you provide. 5. PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels. In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions. [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? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: I Don't Know Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: No Reviewer #2: No ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: The manuscript entitled “Thermostable chaperone-based polypeptide biosynthesis: enfuvirtide model product quality and protocol-related impurities” aimed Enfuvirtide used in HIV infection treatment was synthesized in a developed thermostable chaperone-based peptide biosynthesis system and evaluated for the peptide quality as well as process-related impurities profile. This study is original. There are many grammatical errors in the manuscript. 1. Some references were shown in square brackets, the others were shown in round brackets. For example, square brackets has been used in line 29, while round brackets has been used in line 35. The uniformity should be provided for the manuscript references 2. There are many grammatical errors in the manuscript. These are; -Line 56 – “Thereby, sterilization by filtration is effective only for mycoplasma and virus-free samples”. Thereby should be replaced with the therefore, -Line 58 – “Similarly to chemical synthesis……” should be changed as “Similar to chemical synthesis……” -Line 116 – “Gua 6M with 0.1 M HCl, 0.46 M cyanogen bromide and 4.6% acetonitrile” is an incomplete sentence. -Line 157 - (HPLC PROCESS??) has not been understood. -Line 179 – “IC50 and its 95% confidence interval (CI) estimation was performed using Graphpad prism software (version?, mode)” was should replaced with were. Also, the part “version?” has not been understood. -Line 181 - Graph pad should be written adjacent. -Line 195 – in the last of the sentence “………… according to its manual.,” comma should be removed. -Line 219 – the full stop should removed from the last of the sentence “…….. assessed by HPLC-MS.” -Line 227 – In “The main prodict concentration declined over time”, prodict should be corrected as product. -Line 247 – E. coli should be written italic. -Line 257 to 260 – literature citation should be added. -Reference list format should be checked because it contains some mistakes. For example, Reference 17 includes website address of the publisher and does not include page number. References should be according to the Plos One rules. Reviewer #2: The article entitled “Thermostable chaperone-based polypeptide biosynthesis:enfuvirtide model product quality and protocol-related impurities” by Vladimir Zenin et al, reported about biosynthesis of Enfuvirtide, the therapeutic peptide used in HIV treatment. Biosynthetic methods definitely have few advantages over classical chemical synthesis. The author performed some interesting experiments in-support of their claim; result and discussion were also understandable. Manuscript overall scientific quality is good. However I have some serious doubts regarding their experiments which are listed below. So in my opinion manuscript in the present form the paper is not suitable for publication in Plos one Major Points 1. It’s evident from Figure 1 that the loading of the peptides were not equal in each lane, for e.g. G1 and G16 lane peptides were loaded much lesser compared to FA1 and FA16, they can only quantify digestion if the proteins were loaded in same amount. So in my opinion authors should repeat the experiments with equal loading. 2. Authors reported “significant sample loss for guanidine sample and fusion protein fragmentation for trifluoroacetic acid” however author should provide necessary justification for their observation. 3. Author should also calculate disordered versus ordered structure in Enfuvirtide. It’s not clear to me why chemically manufactured Enfuvirtide is having more disorder? Secondly why their synthesized Enfuvirtide shows similar kind of CD spectra only in 20°C and 65°C why not in other temperatures, is there any disorder to order transition going on with increasing temperature ? In my opinion they should also study peptide conformation in near-UV spectra, this will provide some idea about peptide 3-D structure. 4. It’s not clear to me what is the final yield of Enfuvirtide? Is it comparable to chemical synthesis method or low? 5. Also authors should add a schematic diagram starting from peptide fusion strategy to its final purification Minor Points There are some serious grammatical error, indexing, punctuation and language issue throughout the manuscript so the manuscript needs serious English editing. ****** 6. PLOS authors have the option to publish the peer review history of their article (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 ******** [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.0286752.r001
Revision 1
9 May 2023 Author Response Answers on Reviewer's Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes Reviewer #2: Yes 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: I Don't Know Reviewer #2: Yes 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: No Reviewer #2: No 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: The manuscript entitled “Thermostable chaperone-based polypeptide biosynthesis: enfuvirtide model product quality and protocol-related impurities” aimed Enfuvirtide used in HIV infection treatment was synthesized in a developed thermostable chaperone-based peptide biosynthesis system and evaluated for the peptide quality as well as process-related impurities profile. This study is original. There are many grammatical errors in the manuscript. 1. Some references were shown in square brackets, the others were shown in round brackets. For example, square brackets has been used in line 29, while round brackets has been used in line 35. The uniformity should be provided for the manuscript references Check Zotero results for manuscript (glitched!) 2. There are many grammatical errors in the manuscript. These are; -Line 56 – “Thereby, sterilization by filtration is effective only for mycoplasma and virus-free samples”. Thereby should be replaced with the therefore, (line 48 now, replaced) -Line 58 – “Similarly to chemical synthesis……” should be changed as “Similar to chemical synthesis……” (line 50, changed) -Line 116 – “Gua 6M with 0.1 M HCl, 0.46 M cyanogen bromide and 4.6% acetonitrile” is an incomplete sentence. (corrected, lines 95-98) -Line 157 - (HPLC PROCESS??) has not been understood. (line 127-128 here was link to previous article protocol for me to remember. It is corrected now) -Line 179 – “IC50 and its 95% confidence interval (CI) estimation was performed using Graphpad prism software (version?, mode)” was should replaced with were. Also, the part “version?” has not been understood. (line 145 corrected and replaced by version and build) -Line 181 - Graph pad should be written adjacent. (line 146, corrected) -Line 195 – in the last of the sentence “………… according to its manual.,” comma should be removed. (line 156, removed) -Line 219 – the full stop (dot) should removed from the last of the sentence “…….. assessed by HPLC-MS.” (line 174, removed) -Line 227 – In “The main prodict concentration declined over time”, prodict should be corrected as product. (line 182, corrected) -Line 247 – E. coli should be written italic. line 199, corrected -Line 257 to 260 – literature citation should be added. line 207, I’ve summarized 4 lines properly cited just before, so I had not considered to repeat citation. Now this issue is corrected. -Reference list format should be checked because it contains some mistakes. For example, Reference 17 includes website address of the publisher and does not include page number. References should be according to the Plos One rules. (Had some technical issues, now they are resolved.) Thank you for your comments and expertise, the notes in answer to your comments are in text for your convenience. Reviewer #2: The article entitled “Thermostable chaperone-based polypeptide biosynthesis:enfuvirtide model product quality and protocol-related impurities” by Vladimir Zenin et al, reported about biosynthesis of Enfuvirtide, the therapeutic peptide used in HIV treatment. Biosynthetic methods definitely have few advantages over classical chemical synthesis. The author performed some interesting experiments in-support of their claim; result and discussion were also understandable. Manuscript overall scientific quality is good. However I have some serious doubts regarding their experiments which are listed below. So in my opinion manuscript in the present form the paper is not suitable for publication in Plos one Major Points Thank you for your expertise and engagement. 1. It’s evident from Figure 1 that the loading of the peptides were not equal in each lane, for e.g. G1 and G16 lane peptides were loaded much lesser compared to FA1 and FA16, they can only quantify digestion if the proteins were loaded in same amount. So in my opinion authors should repeat the experiments with equal loading. Theoretically, quantities in samples are the same. In our opinion low recovery and «shadow» on background of G1 could be connected. . We think that the Figure 1 should be represented in such way to highlight low recovery in guanidine for the same initial amount of protein. The final concentration of hydrolysis products is the result we want to highlight. The “substrate”/”products” ratio in final solution is less relevant for process yield than “loaded substrate”/”product in final solution” ratio. Such representation of results is intentional. Sample preparation details were added in Material and Methods section to support and clarify our opinion. 2. Authors reported “significant sample loss for guanidine sample and fusion protein fragmentation for trifluoroacetic acid” however author should provide necessary justification for their observation. Thank you for your comment. In our opinion, the emergence of new lower MW bands on SDS-PAGE of protein sample among with narrowing of main band is a clear sign of fragmentation. (Considering sample simultaneous dilution and contamination with protein material is unrealistic.) We think it is out of article scope to test that shadows and bands by MS methods to prove fragmentation further. 3. Author should also calculate disordered versus ordered structure in Enfuvirtide. It’s not clear to me why chemically manufactured Enfuvirtide is having more disorder? Secondly why their synthesized Enfuvirtide shows similar kind of CD spectra only in 20°C and 65°C why not in other temperatures, is there any disorder to order transition going on with increasing temperature ? In my opinion they should also study peptide conformation in near-UV spectra, this will provide some idea about peptide 3-D structure. DichroWeb structures approximations were added, we changed our statement on disordered structure. Really, only α-helical content is changed substantially from sample to sample. We checked 65°C because same temperature is used for lysate clarification by thermal denaturation of host proteins. While cleavage protocol can be changed to enzymatic, thermal denaturation step is a key feature of our system. So we checked the reversibility of structural processes during heating and revealed such a shift towards original structure. In our opinion, the free dissolved peptide structure (and it is greatly differs from full α-helical in active form on HIV gp41) is less related to peptide activity. We concentrated on discussion of impact of chemical difference on peptide-HIV complex structure. We thoroughly described chemical difference (lines 349-353) and discussed non-acylated C-terminus as the root of structural and antiviral activity difference between biosynthesised and chemically-derived enfuvirtide (lines 354-373). Our discussion was supported by several concordant opinions on C-terminal charge impact on enfuvirtide[1,2] and related peptides[3], and practical decisions[3–6]. We did not analyzed peptides structures in free dissolved form in depth because we used presented structures difference only as demonstration of peptides non-equivalence. We performed NIR spectroscopy, yet our data has variability and our interpretation is limited – peptides are not identical structurally. In our opinion, structural studies are out of scope of present article. 1. Peisajovich SG, Gallo SA, Blumenthal R, Shai Y. C-terminal Octylation Rescues an Inactive T20 Mutant. J Biol Chem. 2003;278: 21012–21017. doi:10.1074/jbc.M212773200 2. Zhang X, Ding X, Zhu Y, Chong H, Cui S, He J, et al. Structural and functional characterization of HIV-1 cell fusion inhibitor T20. AIDS. 2019;33: 1–11. doi:10.1097/QAD.0000000000001979 3. Wild C, Oas T, McDanal C, Bolognesi D, Matthews T. A synthetic peptide inhibitor of human immunodeficiency virus replication: correlation between solution structure and viral inhibition. Proc Natl Acad Sci. 1992;89: 10537–10541. doi:10.1073/pnas.89.21.10537 4. Eckert DM, Kim PS. Design of potent inhibitors of HIV-1 entry from the gp41 N-peptide region. Proc Natl Acad Sci. 2001;98: 11187–11192. doi:10.1073/pnas.201392898 5. Su SB, Gong W, Gao J-L, Shen W-P, Grimm MC, Deng X, et al. T20/DP178, an Ectodomain Peptide of Human Immunodeficiency Virus Type 1 gp41, Is an Activator of Human Phagocyte N-Formyl Peptide Receptor. Blood. 1999;93: 3885–3892. doi:10.1182/blood.V93.11.3885 6. Rimsky LT, Shugars DC, Matthews TJ. Determinants of Human Immunodeficiency Virus Type 1 Resistance to gp41-Derived Inhibitory Peptides. J Virol. 1998;72: 986–993. doi:10.1128/JVI.72.2.986-993.1998 4. It’s not clear to me what is the final yield of Enfuvirtide? Is it comparable to chemical synthesis method or low? The final yield was described in previous article and it is around 35% for lab hydrolysis and HPLC protocol SPPS for enfuvirtide has 6–8% yield for linear process and up to 30% for SPPS and ligation with 7(!) intermediate purification steps for production-scale process(Bray, 2003). Yes, it is comparable in theoretical to practical yield ratio, however the yield per liter of E. coli culture is around 3 mg now. It’s confusing, yet the culture was not even glucose-fed and has potential to increase density more than 10 times. 5. Also authors should add a schematic diagram starting from peptide fusion strategy to its final purification The whole understanding of present article is not possible without previous one. The scheme was introduced in https://doi.org/10.1016/j.btre.2022.e00734 We did not consider to add the process scheme in present article. Yet, now we see how it helps to got the method in a glance, so we added the updated one, thank you. Minor Points There are some serious grammatical error, indexing, punctuation and language issue throughout the manuscript so the manuscript needs serious English editing. We done additional professional grammar check, thank you! 6. PLOS authors have the option to publish the peer review history of their article (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 Attachments Attachment Submitted filename: Answers to rewievers.docx https://doi.org/10.1371/journal.pone.0286752.r002
23 May 2023 Decision Letter - Soumyananda Chakraborti, Editor Thermostable chaperone-based polypeptide biosynthesis: enfuvirtide model product quality and protocol-related impurities PONE-D-22-30362R1 Dear Dr. Zenin, 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 for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. 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, Soumyananda Chakraborti Guest Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0286752.r003
Formally Accepted
31 May 2023 Acceptance Letter - Soumyananda Chakraborti, Editor PONE-D-22-30362R1 Thermostable chaperone-based polypeptide biosynthesis: enfuvirtide model product quality and protocol-related impurities Dear Dr. Zenin: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. 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. If we can help with anything else, please email us at plosone@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. Soumyananda Chakraborti Guest Editor PLOS ONE https://doi.org/10.1371/journal.pone.0286752.r004

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 .