Production of Polyclonal Antibodies and Development of Competitive ELISA for Quantification of the Lantibiotic Paenibacillin

Round 1

Reviewer 1 Report

Title:

 Production of polyclonal antibodies and development of competitive ELISA for quantification of the lantibiotic paenibacillin.

Manuscript ID: 2919048

I recommended that the design of ELISA is very interest and useful. However, for better results of developed method, the screening of a panel of positive and negative samples will be positive. In addition, I would like to know what about crossreactions with others lantibiotic.

Specific suggestions are below:

GENERAL SUGGESTIONS

-          Review scientific names in complete manuscript, including figures and tables. At start a sentence should write complete.

-          Some typographic mistakes and wrong format were appreciated. Format of figures also could be homogenized.

-          Review units according journal instructions.

ABSTRACT

-          In the expression “which is significantly lower tan…” include p value.

MATERIAL AND METHODS

-          Specify number of approbation of ethical committee to use animal.

-          In table 1, authors should specify reference of used primers.

RESULTS

-          In figure 1 and 2, statistical analysis should be included.

-          In figure 3, equation of curve in the figure and correlation will be positive.

-          In figure 5, statistical analysis should be included. Analysis if will be better combine data of panel A and B and panel C with D.

Author Response

Please see the attached file for the detailed response to reviewer's questions

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript describes the development of a competitive ELISA for quantifying the lantibiotic paenibacillin. The work is well organized, accurately describes most methods, and provides important results. However, some adjustments are necessary before being accepted for publication, which are below.

Major points

-It is important to know the molecular weight of the paenibacillin used in the work. This information is important, as this fact can alter the ELISA result when conjugated or not to other molecules.

-Line 118- Please give the constitution (ligand) of the affinity chromatography column.

-Line 123-binding buffer (0.1 M phosphate buffer, 0.15 M NaCl; pH 8); the pH was adjusted using 1 N NaOH. -Please remove the last part of the sentence, as NaOH is not part of the phosphate buffer system, and therefore, the solution will not maintain the desired pH. This buffer must be prepared only with acid and basic phosphate.

-Line 189- remove “Determination of anti-paenibacillin antibody titer” and enter the section number.

-Table 1 - must be inserted as a supplementary

-Line 227- “Figure 1 demonstrates”... Sentence with wrong meaning, change, please.

-Line 451- “exhibited high sensitivity and accuracy”- Inform how much the sensitivity and accuracy is.

-Conclusion- Various repeated information from the introduction is scattered and must be modified, making the conclusions of the study explicit only

-Figure 7 is not well positioned in the conclusions. I suggest placing it in the results or as a Supplement.

The English is reasonably well written; however, it needs a few grammatical revisions and sentence construction.

Author Response

Please see the attached file for the detailed response to reviewer's questions

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

I recommended that complete the number of approbation of ethical committee to use animal in material and method section and include a reference of used primers in supplementary information.

Format of graphic are very different, appear that was carried out by different authors. In my point of view could be homogenized.

Author Response

Please see the attached file

Author Response File: Author Response.docx

Reviewer 2 Report

The authors made an effort to make the manuscript more appropriate. Most of the suggestions were accepted and introduced/restricted in the text, and others were responded to. Therefore, we feel that the manuscript is suitable for acceptance for publication.

Acceptable, and a few grammatical corrections can be made during editing.

Author Response

Thank you for the constructive feedback.