Supramolecular Aggregation Processes on Carbon Surfaces Occurring in Bovine Serum Albumin Solutions
Round 1
Reviewer 1 Report (Previous Reviewer 2)
1 only the image and composition analysis is not enough for publication. some spectra data could be provided.
2. what is the potential applicaiton of your reuslt? and how to apply it.
3. some application related references. please see them. Chinese Chemical Letters 32（2021） 3653-3664; Chinese Chemical Letters 32（2021） 3665-3674
Author Response
Authors are grateful to the Reviewer and appreciate the Reviewer’s suggestions. Authors’ responses are in attached file.
Author Response File: 
Author Response.pdf
Reviewer 2 Report (New Reviewer)
The work entitled “Supramolecular Aggregation Processes on Carbon Surfaces Occurring in Bovine Serum Albumin Solutions” by Aleksei Salanov, Alexandra Serkova, Anastasia Zhirnova, Larisa Perminova and Galina Kovalenko submitted for consideration in MICRO is simple but with proper characterization. It is true that SEM helps, and it is probably the main tool used here. Actually, even though there is no concern about the content of the manuscript, the authors should do more efforts to refer to the most recent work, and it is clear that this part is weak with just 22 references, even though the introduction is not short at all. Take for instance and to expand the scope and interest when discussing the SEM the “sister” technology TEM of transmission electron microscopy could also be included (DOI:10.1016/j.molliq.2022.118675; DOI:10.1002/aoc.6850), and also other uses of SEM (DOI:10.1016/j.jiec.2022.09.026; DOI: 10.1016/j.jcis.2016.11.098), including clearly words like catalysis, including even self-citations (DOI:10.1016/j.clay.2018.03.003) or from other authors (DOI: 10.1002/cctc.202200067)…
Overall, the paper only needs to expand the interest, and once increased the number of references between 35-40 it will be ready for publication.
Author Response
Authors are grateful to the Reviewer. We highly appreciated the Reviewer’s positive attitude towards our manuscript. Authors’ responses arein attached file.
Author Response File: Author Response.pdf
This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.
Round 1
Reviewer 1 Report
The deposition of biological and enzymatically active substances to form thin coatings or film represents an interesting task for many fields as sensors. The manuscript “Supramolecular Aggregation Processes on Carbon Surfaces Occurring in Bovine Serum Albumin Solutions” of Salanov et al. reports a study on aggregation phenomena of bovine serum albumin (BSA) on carbon based supports as HOPG and glassy carbon. Once BSA microdroplets evaporate on support surfaces, protein forms structures that have been characterized through SEM and EDX analysis. The manuscript represents a continuation of a previously published work of the authors (ref. 2) dealing on the protein aggregation on metallic surfaces as aluminum and gold.
While in the previous case, fractal structures have been detected, in this case lacy like replicas are present and located over the entire carbon surface, not just along the defects. Replicas are constituted by bubbles which are clearly visible at higher magnification.
According to the authors, in comparison with metallic surfaces (ref. 2), the formation of fuzzy structures of albumin on carbon surfaces is due to hydrophobic interactions.
This assumption should be more strongly supported by the results, thus a deeper investigation is required.
Moreover, no mention on the pH of BSA solution or characterization data (i.e. surface area) of the supports are reported.
Some relevant points need to be focused.
First four lines are identical to ref. 2.
The authors are strongly recommended to cite qualified and scientific references, thus Wikipedia.org (pg. 4) is not be definitely accepted in any scientific journal.
Although the topic is interesting for scientific community, for all these reasons and mainly for the lack of novelty or relevant improvements, I would not recommend the manuscript for the publication in this journal.
Author Response
The Authors sincerely thank the Reviewer for the remarks, comments, recommendations.
The Authors sincerely thank the Reviewer for the remarks, comments, recommendations.
The authors answer as follows.
Reviewer According to the authors, in comparison with metallic surfaces (ref. 2), the formation of fuzzy structures of albumin on carbon surfaces is due to hydrophobic interactions. This assumption should be more strongly supported by the results; thus, a deeper investigation is required.
Authors agree that " a deeper investigation is required ". On the other hand, there are a lot of studies on the adsorption capacity of carbon materials, e.g. activated carbons, toward various proteins, including albumins, globulins, enzymes. Literature and authors’ data obtained at the MACRO level indicate the presence of the interaction of the protein molecule with the carbon surface (refs. 20, 21) mainly due to hydrophobic Van-der-Waals interactions.
Reviewer Moreover, no mention on the pH of BSA solution or characterization data (i.e. surface area) of the supports are reported.
Authors Albumin solutions were prepared using distilled water with pH 6 ± 0.5 (information was added to text).. Distilled clean water does not contain salts and any impurities; therefore, it was chosen for microscopic studies.
Specific surface area of the HOPG and GC was less than 1 m2/g (information added to text). It is known that the surface of natural graphite is 0.6 m2/g. The surface of glassy carbon (GC) at the MACRO level is dense, smooth and shiny. Glassy carbon crucibles and electrodes are widely used in lab practice.
Since the authors conduct their research at the MICRO (NANO) level, they believe that the macro-characteristics of the supports will not affect the conclusions.
Reviewer First four lines are identical to ref. 2.
Authors try to edit these lines (edited sentence was added to text).
Reviewer The authors are strongly recommended to cite qualified and scientific references, thus Wikipedia.org (pg. 4) is not be definitely accepted in any scientific journal.
Authors agree with the reviewer’ recommendation. We edit this sentence as “Due to the binding of water molecules by one NH2 group of amino acids, a hydrated globule of BSA (600 amino acids) is able to bind thousands of H2O.
Reviewer 2 Report
1. the interaction of material with serum is important to develop new implants. in such a case the background is a little weak. please talk with some Applied Surface Science 308 (2014) 38,,/ Materials Letters 73 (2012) 59–6; references,J. Mater. Chem. A, 2020, 8, 2376, Surface & Coatings Technology 203 (2009) 2017.
2. why you choose carbon as substrate? why not metal? polyers?
3. the identification of the Supramolecular Aggregation Processes should be emphasized .
4. a schematic diagram is required to show the Aggregation Processes you mentioned.
Author Response
The Authors sincerely thank the Reviewer for the remarks, comments, recommendations.
The authors answer are in attached file
Author Response File: Author Response.pdf
Reviewer 3 Report
The presented manuscript shows 20 SEM images of BSA coffee-rings on carbon surfaces. It does not contain any useful information. The self-citation is so high, which is violation of scientific and publication ethics. out of curiosity, I am wondering how authors determined that 80 % of the material formed the coffee-ring?
Author Response
The Authors sincerely thank the Reviewer for the remarks, comments, recommendations.
The Authors sincerely thank the Reviewer for the remarks and questions.
The authors answer as follows.
Reviewer The self-citation is so high, which is violation of scientific and publication ethics.
Authors conduct SEM studies of the aggregation occurring during the drying of microdroplets of albumin solutions deposited on different supports. Comparison of the results obtained here and in previous research (ref.2) is considered justified and expedient by the authors. Apparently, it is this comparison that the Reviewer considers as self-citation. Authors cite their own work in only 2 out of 22 references; self-citation is 9 %. According to the common rules, there is no violation of scientific and publication ethics.
Reviewer: Out of curiosity, I am wondering how authors determined that 80 % of the material formed the coffee-ring?
Authors made a rough qualitative evaluation by calculations, assuming that the edge of the ring (why coffee?) has the shape of a torus cut in half and its volume can be estimated from microscopic images (presented in Fig. 1a,b and in [2]) at the same scaling by elementary measurement with a ruler (in mm). The results of such calculation were listed in Table below.
|
Support |
Albumin concentration, mg·mL–1 |
Relative torus volume units |
|
HOPG |
0.2 |
1.1 |
|
1.0 |
4.7 |
Let's say all the protein molecules are in the “coffee”-ring due to lack of their interaction with the carbon surface. In this case, according to differences in BSA concentration (in 5 times) the volume of the torus should be 5.5 units (not 4.7). Observed difference means that about 15-20% of protein amount was left inside the microdroplet drop, or more than 80% amount of albumin (for HOPG) was in the area of the edge. Undoubtedly, this is a rough estimation, but it also confirms the fact that albumin molecules interact with the carbon surface, in contrast to metallic ones described in first communication (ref.2).
Round 2
Reviewer 1 Report
Although the authors added some additional data in the revised version, as already indicated in the previous revision the manuscript lacks of novelty or "relevant" improvements respect on the related literature and the previous author's manuscript. For these reasons, I cannot recommend the manuscript for the publication.
