Development and characterization of an antibody that recognizes influenza virus N1 neuraminidases

Nan Chen; Renxi Wang; Wanlu Zhu; Xiangjun Hao; Jing Wang; Guojiang Chen; ChunXia Qiao; Xinying Li; Chenghua Liu; Beifen Shen; Jiannan Feng; Lihui Chai; Zuyin Yu; He Xiao

doi:10.1371/journal.pone.0302865

Peer Review History

Original SubmissionDecember 13, 2023
22 Feb 2024 Decision Letter - Victor C Huber, Editor PONE-D-23-41925Development and characterization of an antibody recognizing the influenza virus N1 neuraminidasesPLOS ONE Dear Dr. Xiao, 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. During the revision process, please address comments related to release and presentation of sequence data as well as the comments and/or experiments associated with a challenge study that demonstrates protection against infection. Please submit your revised manuscript by Apr 07 2024 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, Victor C Huber 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. 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. Please see the following video for instructions on linking an ORCID iD to your Editorial Manager account: https://www.youtube.com/watch?v=_xcclfuvtxQ 3. We note that you have included the phrase “data not shown” in your manuscript. Unfortunately, this does not meet our data sharing requirements. PLOS does not permit references to inaccessible data. We require that authors provide all relevant data within the paper, Supporting Information files, or in an acceptable, public repository. Please add a citation to support this phrase or upload the data that corresponds with these findings to a stable repository (such as Figshare or Dryad) and provide and URLs, DOIs, or accession numbers that may be used to access these data. Or, if the data are not a core part of the research being presented in your study, we ask that you remove the phrase that refers to these data. 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. [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: Partly Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes 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: Yes Reviewer #2: Yes ****** 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: This is a straightforward study on the isolation and characterization of human mAb against neuraminidase. Specifically, the mAb appears to be directly targeting N1 as shown in ELISA, cell expression by flow cytometry, and W.B. Furthermore, neuraminidase inhibition assay was performed with finding suggesting rather potent inhibition of enzymatic activities of NA. Moreover, it seems that a.a. residues 219, 254, 358, and 388 in the NA protein were involved in the binding of the viral protien to the mAb. In general, this is a well designed experimental study with data rather clearly being presented. The following points are put forth for the authors to consider: 1. Did the authors test NA subtypes beyond N1, N2 and N7? if not, please explain. Also, a sequence line-up of the NA sequences of the NA used in this study would be helpful for the readerships. 2. While this reviewer does like the work, the manuscript, as it stands now, is quite incomplete in terms of being convincingly showcasing the power of the mAb. Clearly, this work could really be elevated to a much higher visibility by conducting protection experiment in mouse model where mAb is shown to demonstrate protection against chalenge by H1N1, not H3N2 etc. Reviewer #2: In this manuscript, the author got a human cross-reactive H1N1 and H5N1 N1 antibody FNA1 by using phage display. Moreover, this antibody showed NA inhibition and blocking of the virus release and this may be due to steric hinderance. The story is straight-forward and I only have minor comments. 1. For the antibody screened from phage display, is that only one antibody have binding to N1 or have more? 2. The author should submit the FNA1 antibody sequence to public database, like NCBI. 3. For suggestion, the author can use NA protein for ELLA experiments. So the author can test ELLA activity against CA/09 N1. 4. In Figure 3c, the author should describe what is FHA3. In this manuscript, the author obtained a human cross-reactive H1N1 and H5N1 N1 antibody, FNA1, using phage display. Furthermore, this antibody demonstrated NA inhibition and the blocking of virus release, potentially attributable to steric hindrance. The story is straightforward and I only have minor comments. 1. Is the antibody screened from phage display the only one binding to N1, or are there others? 2. The author should consider submitting the FNA1 antibody sequence to public databases like NCBI. 3. As a suggestion, the author could use NA protein for ELLA experiments to assess ELLA activity against CA/09 N1. 4. In Figure 3c, the author should clarify the nature of FHA3. ****** 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.0302865.r001
Revision 1
27 Mar 2024 Author Response Response to academic editor 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 Response: In accordance with PLOS ONE's style requirements, we have adjusted the formatting of our manuscript entitled “Development and characterization of an antibody recognizing the influenza virus N1 neuraminidases” and followed the file naming process. 2. 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. Please see the following video for instructions on linking an ORCID iD to your Editorial Manager account: https://www.youtube.com/watch?v=_xcclfuvtxQ Response: ORCID iDs of the three corresponding authors of this manuscript have been successfully linked to their Editorial Manager accounts following the provided instructions. 3. We note that you have included the phrase “data not shown” in your manuscript. Unfortunately, this does not meet our data sharing requirements. PLOS does not permit references to inaccessible data. We require that authors provide all relevant data within the paper, Supporting Information files, or in an acceptable, public repository. Please add a citation to support this phrase or upload the data that corresponds with these findings to a stable repository (such as Figshare or Dryad) and provide and URLs, DOIs, or accession numbers that may be used to access these data. Or, if the data are not a core part of the research being presented in your study, we ask that you remove the phrase that refers to these data. Response: We acknowledge this suggestion and have added tables in the supporting information (S2 and S3 Tables) to confirm our statement. 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. Response: We have updated the supporting information files by replacing the original ones with a Word file (S1_File.docx), a figure (S1_Fig.png), and three Excel files (S1_Table.xlsx, S2_Table.xlsx, and S3_Table.xlsx). In addition, captions for the supporting information files have been added at the end of our manuscript, following the guidelines for supporting information. We have also updated the in-text citations accordingly (line 292, line 306, and line 330). Response to Reviewer #1 1. Did the authors test NA subtypes beyond N1, N2 and N7? if not, please explain. Also, a sequence line-up of the NA sequences of the NA used in this study would be helpful for the readerships. Response: We focused our testing exclusively on NA subtypes N1 (CA/09 NA, PR8 NA, BR/07 NA, and Anh05 NA), N2 (H3N2 NA), and N9 (H7N9 NA). The reason for this is that CA/09 NA was used as the target antigen during phage display selections. Therefore, the antibodies obtained are probably N1-specific. While FNA1 demonstrated binding to CA/09 and Anh05 NA, it exhibited no reactivity with PR8, BR/07, H3N2, and H7N9 NA, which shows its limited binding breadth. Additionally, in response to the reviewer’s recommendation, we aligned the NA amino acid sequences of influenza viruses described in the study, and found that Anh05 NA showed a higher homology between Anh05 NA and CA/09 NA compared to other sequences (line 290–292, S1 Fig and S1 Table). 2. While this reviewer does like the work, the manuscript, as it stands now, is quite incomplete in terms of being convincingly showcasing the power of the mAb. Clearly, this work could really be elevated to a much higher visibility by conducting protection experiment in mouse model where mAb is shown to demonstrate protection against challenge by H1N1, not H3N2 etc. Response: We acknowledge the reviewer’s suggestion regarding mouse challenge experiments in terms of demonstrating the protective capacity of the mAb against H1N1, but not other subtypes, e.g., H3N2, which could significantly improve the visibility of our study. Regrettably, we currently lack access to the CA/09 virus. Nevertheless, we carried out an in vivo prophylactic study to evaluate FNA1 mAb's potential in protecting against PR8. Despite our efforts, FNA1 failed to provide protection against PR8, as shown in the subsequent figure. Response to Reviewer #2 1. Is the antibody screened from phage display the only one binding to N1, or are there others? Response: Through the use of phage display techniques, we identified three mAbs targeting N1 NAs. While all three mAbs showed binding activity to CA/09 and Anh05 NA, the affinities (KD) of the two other mAbs were lower compared to FNA1. In addition, analysis revealed that these three mAbs shared the same four key residues of H1N1 NA that affected their binding to viral proteins, suggesting that they might target similar sites. Based on these findings, we opted to focus solely on FNA1 for presenting the results. 2. The author should consider submitting the FNA1 antibody sequence to public databases like NCBI. Response: We concur with the reviewer’s suggestion regarding the submission of the FNA1 mAb sequence to a public database. Consequently, we have deposited the DNA sequences of both the FNA1 heavy and light chain variable regions at GenBank. The accession numbers for the nucleotide sequences are PP483115 (heavy chain) and PP483116 (light chain). 3. As a suggestion, the author could use NA protein for ELLA experiments to assess ELLA activity against CA/09 N1. Response: We agree with the reviewer’s suggestion to utilize NA protein in ELLA experiments to assess the NI activity of FNA1 mAb against CA/09 N1. Accordingly, we conducted ELLA assays using recombinant CA/09 NA and Anh05 NA, and found that FNA1 effectively blocked the activity of both CA/09 NA and Anh05 NA, resulting in the decreased cleavage of fetuin by NA proteins, with IC50 values of 0.06 μg/mL and 2.88 μg/mL, respectively (line 306–309 and updated Fig 3b). 4. In Figure 3c, the author should clarify the nature of FHA3. Response: We greatly appreciate the reviewer's suggestion and have added a clarifying remark in line 319. Attachments Attachment Submitted filename: Response to Reviewers(2).docx https://doi.org/10.1371/journal.pone.0302865.r002
4 Apr 2024 Decision Letter - Victor C Huber, Editor PONE-D-23-41925R1Development and characterization of an antibody that recognizes influenza virus N1 neuraminidasesPLOS ONE Dear Dr. Xiao, 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. During the revision process, please clearly state and expand on the limitations of the study. Please submit your revised manuscript by May 19 2024 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, Victor C Huber Academic Editor PLOS ONE Journal Requirements: Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: (No Response) ******** 2. 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: Partly ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: I Don't Know ****** 4. 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 ****** 5. 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: Yes ****** 6. 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: Please see my responses to your revision Ms: Response to Reviewer #1 1. Did the authors test NA subtypes beyond N1, N2 and N7? if not, please explain. Also, a sequence line-up of the NA sequences of the NA used in this study would be helpful for the readerships. Response: We focused our testing exclusively on NA subtypes N1 (CA/09 NA, PR8 NA, BR/07 NA, and Anh05 NA), N2 (H3N2 NA), and N9 (H7N9 NA). The reason for this is that CA/09 NA was used as the target antigen during phage display selections. Therefore, the antibodies obtained are probably N1-specific. While FNA1 demonstrated binding to CA/09 and Anh05 NA, it exhibited no reactivity with PR8, BR/07, H3N2, and H7N9 NA, which shows its limited binding breadth. Additionally, in response to the reviewer’s recommendation, we aligned the NA amino acid sequences of influenza viruses described in the study, and found that Anh05 NA showed a higher homology between Anh05 NA and CA/09 NA compared to other sequences (line 290–292, S1 Fig and S1 Table). Reviewer's responses to the latest revised Ms: Understood your limitations. Yet, such limitations should be more elaborated in the Discussion. 2. While this reviewer does like the work, the manuscript, as it stands now, is quite incomplete in terms of being convincingly showcasing the power of the mAb. Clearly, this work could really be elevated to a much higher visibility by conducting protection experiment in mouse model where mAb is shown to demonstrate protection against challenge by H1N1, not H3N2 etc. Response: We acknowledge the reviewer’s suggestion regarding mouse challenge experiments in terms of demonstrating the protective capacity of the mAb against H1N1, but not other subtypes, e.g., H3N2, which could significantly improve the visibility of our study. Regrettably, we currently lack access to the CA/09 virus. Nevertheless, we carried out an in vivo prophylactic study to evaluate FNA1 mAb's potential in protecting against PR8. Despite our efforts, FNA1 failed to provide protection against PR8, as shown in the subsequent figure. Reviewer's responses to the latest revised Ms: Understood your limitations. Yet, such limitations should be more clearly elaborated in the Discussion. ****** 7. 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 ******** [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.0302865.r003
Revision 2
11 Apr 2024 Author Response Article Title: Development and characterization of an antibody recognizing the influenza virus N1 neuraminidases Running Number: PONE-D-23-41925R1 Response to academic editor 1. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. Response: We acknowledge this suggestion and have reviewed the reference list carefully. We discovered that the 30th reference was incomplete, as it lacked the identifier (DOI), and the 31st reference was a manuscript that had just been accepted. Consequently, these two references were removed and replaced with relevant current references (line 499–504). Response to Reviewer #1 1. Did the authors test NA subtypes beyond N1, N2 and N7? if not, please explain. Also, a sequence line-up of the NA sequences of the NA used in this study would be helpful for the readerships. Response: We focused our testing exclusively on NA subtypes N1 (CA/09 NA, PR8 NA, BR/07 NA, and Anh05 NA), N2 (H3N2 NA), and N9 (H7N9 NA). The reason for this is that CA/09 NA was used as the target antigen during phage display selections. Therefore, the antibodies obtained are probably N1-specific. While FNA1 demonstrated binding to CA/09 and Anh05 NA, it exhibited no reactivity with PR8, BR/07, H3N2, and H7N9 NA, which shows its limited binding breadth. Additionally, in response to the reviewer’s recommendation, we aligned the NA amino acid sequences of influenza viruses described in the study, and found that Anh05 NA showed a higher homology between Anh05 NA and CA/09 NA compared to other sequences (line 290–292, S1 Fig and S1 Table). Reviewer's responses to the latest revised Ms: Understood your limitations. Yet, such limitations should be more elaborated in the Discussion. Response: We concur with the reviewer’s suggestion regarding the exposition of the limitations of our study in the Discussion, and we have added further relevant content to clarify the limitations (line 370–374). 2. While this reviewer does like the work, the manuscript, as it stands now, is quite incomplete in terms of being convincingly showcasing the power of the mAb. Clearly, this work could really be elevated to a much higher visibility by conducting protection experiment in mouse model where mAb is shown to demonstrate protection against challenge by H1N1, not H3N2 etc. Response: We acknowledge the reviewer’s suggestion regarding mouse challenge experiments in terms of demonstrating the protective capacity of the mAb against H1N1, but not other subtypes, e.g., H3N2, which could significantly improve the visibility of our study. Regrettably, we currently lack access to the CA/09 virus. Nevertheless, we carried out an in vivo prophylactic study to evaluate FNA1 mAb's potential in protecting against PR8. Despite our efforts, FNA1 failed to provide protection against PR8, as shown in the subsequent figure. Reviewer's responses to the latest revised Ms: Understood your limitations. Yet, such limitations should be more clearly elaborated in the Discussion. Response: We greatly appreciate the reviewer's suggestion and have clearly stated the limitations in the Discussion (line 400–404). Attachments Attachment Submitted filename: Response to Reviewers(2).docx https://doi.org/10.1371/journal.pone.0302865.r004
15 Apr 2024 Decision Letter - Victor C Huber, Editor Development and characterization of an antibody that recognizes influenza virus N1 neuraminidases PONE-D-23-41925R2 Dear Dr. Xiao, 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. If you have any questions relating to publication charges, 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, Victor C Huber Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0302865.r005
Formally Accepted
29 Apr 2024 Acceptance Letter - Victor C Huber, Editor PONE-D-23-41925R2 PLOS ONE Dear Dr. Xiao, 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 If revisions are needed, the production department will contact you directly to resolve them. If no revisions are needed, you will receive an email when the publication date has been set. At this time, we do not offer pre-publication proofs to authors during production of the accepted work. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few weeks 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. 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. Victor C Huber Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0302865.r006

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