N-Glycosylation Profile of Abrin Certified EU Reference Material

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript describing the glycosylation pattern for the abrin certified reference material developed by the EuroBioTox consortium is generally well-written and the figures/tables presented support the conclusions of the authors. The fact that a certified abrin reference that is well-characterized is a significant accomplishment especially to those that work on these plant toxins.

The manuscript is ready for publication after some minor revisions/comments to make it easier for the general audience to understand better:

a)Figure 1:  Suggest that the Y-axis light blue color be changed to something easier to read (maybe bold and/or another solid color that is easier to distinguish). Additionally, the three color traces are hard to distinguish from each other especially with blue/green- maybe revise using something with more contrast. 

b)Figure 3: Suggest add a description of what F (M2FX) means in the figure legend. It is described in line 119 but addition to figure legend will allow readers to quickly know what they are looking at.

c)Line 120: There seems to be a duplication of ", refer to Table 2 Table2for structures". Please edit accordingly.

d)Figure 5: Y-axis states A/%-  For the general audience, maybe suggest Y-axis should be labelled : Relative amounts [Area/%]?

e)If change to Y-axis figure 5, then Table 2 A/% should be changed too

f) Figure 6: Please add a little more detail/conclusion(main point) to the figure legend. It is a bit brief.

g) Line 345/: ", allegedly is no requirement for ..." Do you mena , allegedly is not required for protein integrity....?

 

 

Author Response

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Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript did a lot of work, however, there are some questions and suggestions list below:

1. Line 377~379: explain the reasons for changing the type of added acid.

2. Adjust the sequence of the Results part and Materials and Methods part.

3. Fig.4(a): Explain the different connotations of the structure of peaks simultaneously in M8 and M7.

4. Fig.9(a): Don't overlap words.

5. Fig.9(c)-7: Text layout should not exceed the picture.

6. Line 66: Whether the buffer without galactose interference can be selected?

Comments on the Quality of English Language

The language needs to be improved.

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The paper describes in great detail the N-Glycosylation profile of certified Abrin EU reference material. The paper is well presented and as expected goes into much detail about the  N-Glycosylation profile of Abrin. 

I believe the paper is well written and executed, the results and methods are all well presented and the paper warrents publication. 

There are only two point I would make for the improvement of the paper:

Firstly on Lines 20-22 and lines 51-53 the authors say that the N-glycosylation profile could be used to determine sample provenance, I think this could be a really nice discussion point and I would ask the author to perhap add more to the discussion, talking about how this profile they have defined could be used in that way. 

Secondly and it is a shame that the authors did not also perform the same analysis on a second batch of uncertified Abrin to compare between two different batches. This would have highlighted any potential area of different that could aid in my point above.  

Neither of the two above comments distract from what is a nice high quality piece of science that should be published. 

 

Author Response

Please see the attachment

Author Response File: Author Response.pdf