Dear Dr. Luna,

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.

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We look forward to receiving your revised manuscript.

Kind regards,

Satish kumar Rajasekharan

Academic Editor

PLOS ONE

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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

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

Reviewer #1: Yes

Reviewer #2: Yes

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

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: The manuscript presents a well-designed and scientifically sound study addressing the urgent issue of antimicrobial resistance, focusing on Acinetobacter baumannii and Pseudomonas aeruginosa. The exploration of monoclonal antibodies as an alternative to small-molecule therapeutics is both timely and innovative. The study is clearly written in standard scientific English, with a logical structure and coherent flow of ideas.The authors have made all data underlying the findings fully available, demonstrating transparency and commitment to reproducibility. The inclusion of both in vitro assays and in vivo infection models strengthens the reliability of the results. Statistical analyses appear to have been performed appropriately and rigorously, supporting the validity of the conclusions.The findings are technically sound and well interpreted. The distinction between in vitro cross-reactivity and the lack of in vivo efficacy against P. aeruginosa is handled with scientific caution, and the discussion appropriately links these observations to the broader goal of developing broad-spectrum therapeutic antibodies.Overall, the manuscript is methodologically robust, well written, and contributes valuable insights into antibody-based strategies against multidrug-resistant pathogens. I recommend publication after minor editorial revisions.

Reviewer #2: The manuscript primarily reports cross-reactivity of a previously characterized antibody; however, the novelty is limited since the lack of in vivo efficacy diminishes translational impact. Authors should better justify the originality of this work relative to prior studies

The rationale for testing P. aeruginosa appears speculative. A more systematic antigenic or epitope-level comparison should precede experimental testing to justify this direction.

The central hypothesis—that MAb10 binds pseudaminic acid-containing epitopes across species—requires biochemical validation. Without structural confirmation (e.g., glycan array or ELISA with purified carbohydrates), this remains speculative. Also, authors must discuss alternative explanations for cross-reactivity, such as nonspecific binding or shared glyco-motifs unrelated to pseudaminic acid.

The in vitro binding results rely solely on flow cytometry. There is no orthogonal confirmation (e.g., ELISA, immunoblot, or surface plasmon resonance).

The use of only one or two P. aeruginosa reference strains (O7 and O9) limits generalizability. Broader strain diversity should be analyzed or statistically summarized.

The opsonophagocytosis assay shows marginal significance (p = 0.0476) and a modest biological effect. Replication and quantitative statistical reporting (effect size, SD, n) are needed.

The manuscript lacks proper negative and positive controls for cross-reactivity—e.g., known P. aeruginosa O-antigen MAb as a comparator.

Antibody purity, concentration verification, and endotoxin levels should be described since these can affect macrophage responses and animal outcomes.

There is no mention of antibody affinity or subclass, both of which influence effector function and could explain the lack of protection.

The manuscript presents a study on cross-species reactivity of an anti-A. baumannii monoclonal antibody (MAb10) against Pseudomonas aeruginosa. The topic is timely and relevant, considering the global concern over antimicrobial resistance. However, while the study demonstrates some interesting in vitro observations, the experimental depth, mechanistic exploration, and translational significance are limited. The work lacks sufficient novelty and mechanistic insights to warrant publication in its current form. This manuscript would likely require major revisions before it could meet PLOS ONE standards.

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Reviewer #1: No

Reviewer #2: Yes:  Chaitany Jayprakash Raorane

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To ensure your figures meet our technical requirements, please review our figure guidelines: https://journals.plos.org/plosone/s/figures 

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A monoclonal antibody raised against Acinetobacter baumannii capsular carbohydrate exhibits cross-species in vitro binding against Pseudomonas aeruginosa

PONE-D-25-55530R1

Dear Dr. Luna,

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.

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Kind regards,

Satish kumar Rajasekharan

Academic Editor

PLOS One